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  • 77
    Millipore p21 antigen extraction agent
    The effect of pifithrin-α on <t>p21</t> levels of hepatocytes in the presence of HGF. Rat hepatocytes were cultured at high density in WE containing 10% FCS, 10 ng/mL HGF, along with various concentrations of pifithrin-α, the chemical inhibitor of p53, dissolved in DMSO, or DMSO of the same concentration for 18 hours. An open bar denotes hepatocytes cultured in the absence of HGF. Closed bars denote hepatocytes cultured with pifithrin-α in the presence of 10 ng/mL HGF. Dotted bars denote hepatocytes cultured with DMSO in the presence of 10 ng/mL HGF. Data are mean ± SEM of four dishes. *p
    P21 Antigen Extraction Agent, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p21 antigen extraction agent/product/Millipore
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    p21 antigen extraction agent - by Bioz Stars, 2019-05
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    93
    GE Healthcare contrast agent iohexol
    Survival of RC rats receiving single IV 2.5 g I/kg doses of <t>iohexol</t> or iohexol + SBECD (sulfobutyl‐either β cyclodextrin) at mole ratio of 1:0.025 iohexol:SBECD. N = 8 in each group.
    Contrast Agent Iohexol, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/contrast agent iohexol/product/GE Healthcare
    Average 93 stars, based on 20 article reviews
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    contrast agent iohexol - by Bioz Stars, 2019-05
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    90
    Selleck Chemicals antitumor agent sorafenib
    Copine-III regulated the sensitivity of cells to <t>sorafenib.</t> Notes: (A) MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with indicated concentrations of sorafenib and analyzed by MTT assays. ( B ) MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with IC 50 concentration of sorafenib and analyzed by MTT assays at indicated time points. ( C ) L-02 cells, which were infected with control or CPNE3, were treated with indicated concentration of sorafenib and analyzed by MTT-assays. ( D ) MHCC97-H cells, which were infected with control or CPNE3, were treated with the IC 50 concentration of sorafenib and analyzed by MTT assays at indicated time points. * P
    Antitumor Agent Sorafenib, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 90/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 90 stars, based on 18 article reviews
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    antitumor agent sorafenib - by Bioz Stars, 2019-05
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    82
    Guerbet contrast agent endorem
    Prussian Blue staining of human chondrocytes labeled with ( A ) <t>Endorem</t> ® (Guerbet, Roissy, France), ( B ) dopamine-hyaluronate-maghemite nanoparticles Run IIIB/3, ( C ) hyaluronate-maghemite nanoparticles Run II/3, and ( D ) dopamine-hyaluronate-maghemite nanoparticles Run IIIC/3. Notes: Cell nuclei are counterstained with nuclear fast red. Scale bar 25 μm.
    Contrast Agent Endorem, supplied by Guerbet, used in various techniques. Bioz Stars score: 82/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Millipore denitrosylating agent
    S-nitrosylation of eNOS is increased in the penis after cavernous nerve injury, and is decreased by treatment with a <t>denitrosylating</t> agent NAC. eNOS S-nitrosylation was measured by TMT-switch assay consisting of immunoprecipitation of TMT-tagged proteins with anti-eNOS antibody, followed by Western blot against TMT. Upper panels are representative Western immunoblots of eNOS-SNO and β-actin in penes of Sham+Vehicle, Sham+NAC, BCNI+Vehicle, and BCNI+NAC rats (lanes 2–5). Lower panel represents quantitative analysis of eNOS-SNO/β-actin in the same treatment groups. No S-nitrosylation of eNOS could be detected in penile tissue of BCNI+Vehicle rats in the absence of ascorbate (lane 1). n=3
    Denitrosylating Agent, supplied by Millipore, used in various techniques. Bioz Stars score: 80/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Several laboratories Inc therapeutic agents targeting pkcα
    Disrupting the interaction between MZF-1 and Elk-1 decreases EMT potential ( A ) Comparison of the gene expression profiles of upregulated (left panel) and downregulated (right panel) EMT-related genes in MZF-1 60–72 -transfected Hs578T and MB-231 cloned cells, as determined by microarray with those of the parental cells. ( B ) Immunoblotting analysis of changes in protein levels in the parental and transfected cells and in the less malignant MB-468 (MB-468) and MCF-7 cells. ( C ) Visualization and quantification of cell migration of <t>PKCα-transfected</t> cells by migration assay. MZF-1 60–72 construct-transfected stably cloned cells were transfected with the empty-vector or full-length PKCα construct for 3 days, and migration assay was performed. ** p
    Therapeutic Agents Targeting Pkcα, supplied by Several laboratories Inc, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Chimerigen Laboratories costimulatory blockade agents
    A : Porcine insulin levels in the serum of C57BL mice transplanted with E42 pancreas and treated with <t>costimulatory</t> blockade agents (anti-LFA1, anti-CD48, and ±CTLA4-Ig), FTY720, with (◇) or without (○) debulking, at different time points after transplantation. Treatment with costimulatory antibodies was stopped at 3 months posttransplant, and graft maintenance was continued twice weekly only with FTY720. Insulin levels in the serum of NOD-SCID mice transplanted with E42 pancreas served as a positive control (♦). The inset demonstrates average pig insulin levels in transplanted mice over a course of 6 months. No statistical difference could be found between the tested groups. B : Porcine insulin levels in the serum of C57BL mice transplanted with E42 pancreas and treated with costimulatory blockade agents with or without debulking at different time points after FTY720 withdrawal. Data are presented as means ± SE.
    Costimulatory Blockade Agents, supplied by Chimerigen Laboratories, used in various techniques. Bioz Stars score: 78/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/costimulatory blockade agents/product/Chimerigen Laboratories
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    78
    Millipore agent cisplatin
    Repeated systemic injections of URB597 (0.3 mg/kg daily, i.p.) reduced the development of <t>cisplatin-evoked</t> hyperalgesia in cisplatin-treated mice. A ) When cisplatin was co-injected with URB597, the development of mechanical hyperalgesia was delayed, and
    Agent Cisplatin, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Nanoprobes metal nanoparticle contrast agents
    Dual energy micro-CT material decomposition. (A) , In vitro phantom consisting of a large tube of water surrounded by vials containing gold, iodine, or a mixture of the two. (B) , In vivo imaging of gold <t>nanoparticles</t> and iodine-containing liposomes within a mouse soft tissue sarcoma. The iodine (shown in red) and gold (shown in green) maps are the result of dual energy decomposition. In both cases, the decomposition was able to successfully differentiate the signals from the gold and iodine contrast agents.
    Metal Nanoparticle Contrast Agents, supplied by Nanoprobes, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    R&D Systems u3 1287 agents
    Dual blockade of HER3 and EGFR can effectively inhibit the proliferation of Ctx R clones. (A) Combinatorial treatment of Ctx R clones with cetuximab and <t>U3-1287</t> leads to proliferation inhibition. Cell proliferation was measured using crystal violet assay and plotted as a percentage of proliferation relative to the vehicle control cells. Data points are represented as mean ± s.e.m. (n = 3). (B) Combinatorial treatment with cetuximab and U3-1287 leads to loss of HER3 expression in Ctx R clones. Protein lysates were fractionated on SDS–PAGE followed by immunoblotting for the indicated proteins. α-Tubulin was used as a loading control.
    U3 1287 Agents, supplied by R&D Systems, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore trpv1 specific agent capsaicin
    Elevated IOP increases <t>TRPV1</t> in DBA/2 mice. Immunolabeling against TRPV1 with DAPI counterstain in retina from DBA/2 mice ( A, B ) and colabeling of the RGC-specific marker Thy1 and TRPV1 C57 retina for comparison ( C–E ). All micrographs from the
    Trpv1 Specific Agent Capsaicin, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    XL-protein tivantinib single agent
    Identification of <t>tivantinib</t> and ABT-199 as a synergistic drug combination in AML cells. ( a ) Results of tivantinib combination drug screen in HL60 cells using a customized library of 240 targeted agents. Replicate correlations of cell viability following treatment with individual library compounds (2.5 μM) (left) and compounds in combination with tivantinib (0.25 μM) (middle) are displayed. Fold change corresponds to the ratio of inhibition of cell viability achieved by a drug combination with tivantinib (0.25 μM) compared to individual single library compounds (2.5 μM). Drugs passing fold change > 1.5 cutoff are highlighted in red. Navitoclax and ABT-199 are labeled. ( b ) Dose response curves for inhibition of viability of HL60 cells of tivantinib and its combination with either ABT-199 (left) or cytarabine (Ara-C; right). Synergy is assessed by the Bliss model of independence (histograms in insets). Displayed in the histograms are the experimentally determined differences for each drug combination from the calculated Bliss additivity on a scale of +20% to −20% cell viability in order of increasing tivantinib concentrations. Vertical lines indicate increments of 10% cell viability. Bars pointing up from the blue baseline (additivity) indicate synergy, bars pointing down indicate antagonism. ( c ) Effects of tivantinib and ABT-199 combination (in μM) on PARP-1 and caspase 3 cleavage as well as pSer10 histone H3 levels after 24 h treatment. ( d ) Effects of tivantinib and ABT-199 combination on β-catenin stabilization and pGSK3α/β Y279/216 levels. ( e ) Effects of tivantinib and ABT-199 combination on MCL-1, BCL-XL, and Bak. V = vehicle (DMSO). Tivantinib, ABT-199, and BIO concentrations are in μM. NaCl and LiCl concentrations are in mM.
    Tivantinib Single Agent, supplied by XL-protein, used in various techniques. Bioz Stars score: 76/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Syndax sndx 275 agent
    The effect of treatment with <t>SNDX-275</t> on Fas and FADD expression in LM7 lung metastases in vivo
    Sndx 275 Agent, supplied by Syndax, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    ProSense cathepsin activated agent prosense
    Immunofluorescence histochemistry of the sections from the aortic arch apoE −/− mice on a high cholesterol diet. (a) IntegriSense, (b) <t>ProSense,</t> red: imaging agent, green: CD 68 antibody staining monocytes/macrophages, blue: DAPI staining of cell nucleus, yellow arrows: colocalization of agents and macrophages, red arrows: distribution of agents in the core of atherosclerotic lesion, Adv: adventitia.
    Cathepsin Activated Agent Prosense, supplied by ProSense, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Gilead Sciences agents ambisome
    In vitro killing assay. Notes: Intracellular antifungal activity of the AmB–PGA formulation against Candida albicans inside RAW 264.7 cells. Macrophages were infected with C. albicans and treated with AmB–PGA nanoparticles. Percentage of infection was calculated by lysing macrophages (after 24 hours of treatment) with Triton X-100 (0.2%), subculturing in Sabouraud agar plates, and comparing with an untreated control. Experiments were carried out in triplicate. Each datum point represents the mean ± standard deviation. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, <t>Ambisome</t> ® ; CFU, colony forming units; PGA, polyglutamic acid.
    Agents Ambisome, supplied by Gilead Sciences, used in various techniques. Bioz Stars score: 76/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Millipore lipid raft disrupting agents mβcd
    Effect of <t>MβCD</t> on cell cycle and apoptosis of triple negative breast cancer cells. Cell cycle distribution of MDA-MB 231 (A) and MDA-MB 468 cells (B). Propidium iodide stained cells were analyzed for DNA content using flow cytometry. Histograms represent the percentage of MDA-MB 231 (C) and MDA-MB 468 (D) cells in G0/G1, S and G2/M phases. The data represent one of three independent experiments. Values are mean±SD of three different experiments. MDA-MB 231 (E) and MDA-MB 468 (F) cells were stained for apoptosis using TdT-mediated dUTP nick end-labeling (TUNEL) assay. Quantification of apoptotic cells expressed as a percent of 4′,6-diamidino-2-phenylindole (DAPI)-stained cells in MDA-MB231 (G) and MDA-MB 468 (H). Data shown from three independent experiments, bars represent the mean±SD of three experiments.
    Lipid Raft Disrupting Agents Mβcd, supplied by Millipore, used in various techniques. Bioz Stars score: 76/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Advanced Cancer Therapeutics Inc single agent motolimod
    Preclinical studies of <t>motolimod</t> (Moto) plus pegylated liposomal doxorubicin (PLD) in NSG-HIS mice. (A) NSG-HIS mice were given either PLD alone (intraperitoneally, 50 mg/m 2 ), motolimod alone (subcutaneously 2 days after PLD dosing, 1.5 mg/m 2 ), the combination,
    Single Agent Motolimod, supplied by Advanced Cancer Therapeutics Inc, used in various techniques. Bioz Stars score: 76/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    75
    Millipore cuo nps agent
    Phases and morphologies of the solid products from reaction of <t>CuO</t> <t>NPs</t> with soluble HS - . (A) XRD patterns of sulfidated CuO nanoparticles generated from initial Cu/S ratios that vary from 0.2 to 5. CuO (tenorite) and CuS (covellite) reference is presented
    Cuo Nps Agent, supplied by Millipore, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The effect of pifithrin-α on p21 levels of hepatocytes in the presence of HGF. Rat hepatocytes were cultured at high density in WE containing 10% FCS, 10 ng/mL HGF, along with various concentrations of pifithrin-α, the chemical inhibitor of p53, dissolved in DMSO, or DMSO of the same concentration for 18 hours. An open bar denotes hepatocytes cultured in the absence of HGF. Closed bars denote hepatocytes cultured with pifithrin-α in the presence of 10 ng/mL HGF. Dotted bars denote hepatocytes cultured with DMSO in the presence of 10 ng/mL HGF. Data are mean ± SEM of four dishes. *p

    Journal: PLoS ONE

    Article Title: Induction of p53-Dependent p21 Limits Proliferative Activity of Rat Hepatocytes in the Presence of Hepatocyte Growth Factor

    doi: 10.1371/journal.pone.0078346

    Figure Lengend Snippet: The effect of pifithrin-α on p21 levels of hepatocytes in the presence of HGF. Rat hepatocytes were cultured at high density in WE containing 10% FCS, 10 ng/mL HGF, along with various concentrations of pifithrin-α, the chemical inhibitor of p53, dissolved in DMSO, or DMSO of the same concentration for 18 hours. An open bar denotes hepatocytes cultured in the absence of HGF. Closed bars denote hepatocytes cultured with pifithrin-α in the presence of 10 ng/mL HGF. Dotted bars denote hepatocytes cultured with DMSO in the presence of 10 ng/mL HGF. Data are mean ± SEM of four dishes. *p

    Article Snippet: The suspensions were incubated with p21 antigen extraction agent (1.0 M potassium chloride, 6% zwittergent (Calbiochem, CA)), and centrifuged.

    Techniques: Cell Culture, Concentration Assay

    Cellular p21 levels in hepatocytes treated with HGF. Rat hepatocytes were cultured in WE containing 10% FCS and various concentration of HGF for 18 hours. Closed bars denote hepatocytes cultured at high density. Open bars denote hepatocytes cultured at low density. Data are mean + SEM of four dishes. *p

    Journal: PLoS ONE

    Article Title: Induction of p53-Dependent p21 Limits Proliferative Activity of Rat Hepatocytes in the Presence of Hepatocyte Growth Factor

    doi: 10.1371/journal.pone.0078346

    Figure Lengend Snippet: Cellular p21 levels in hepatocytes treated with HGF. Rat hepatocytes were cultured in WE containing 10% FCS and various concentration of HGF for 18 hours. Closed bars denote hepatocytes cultured at high density. Open bars denote hepatocytes cultured at low density. Data are mean + SEM of four dishes. *p

    Article Snippet: The suspensions were incubated with p21 antigen extraction agent (1.0 M potassium chloride, 6% zwittergent (Calbiochem, CA)), and centrifuged.

    Techniques: Cell Culture, Concentration Assay

    Cellular p53 and p21 levels in hepatocytes treated with HGF. Rat hepatocytes were cultured in WE containing 10% FCS and various concentrations of HGF for 18 hours. Open circles denote hepatocytes cultured in the absence of HGF. Dotted open circles denote hepatocytes cultured with 2.5 ng/mL HGF. Dotted closed circles denote hepatocytes cultured with 5 ng/mL HGF. Closed circles denote hepatocytes cultured with 10 ng/mL HGF. (A) Hepatocytes cultured at high density. (B) Hepatocytes cultured at low density.

    Journal: PLoS ONE

    Article Title: Induction of p53-Dependent p21 Limits Proliferative Activity of Rat Hepatocytes in the Presence of Hepatocyte Growth Factor

    doi: 10.1371/journal.pone.0078346

    Figure Lengend Snippet: Cellular p53 and p21 levels in hepatocytes treated with HGF. Rat hepatocytes were cultured in WE containing 10% FCS and various concentrations of HGF for 18 hours. Open circles denote hepatocytes cultured in the absence of HGF. Dotted open circles denote hepatocytes cultured with 2.5 ng/mL HGF. Dotted closed circles denote hepatocytes cultured with 5 ng/mL HGF. Closed circles denote hepatocytes cultured with 10 ng/mL HGF. (A) Hepatocytes cultured at high density. (B) Hepatocytes cultured at low density.

    Article Snippet: The suspensions were incubated with p21 antigen extraction agent (1.0 M potassium chloride, 6% zwittergent (Calbiochem, CA)), and centrifuged.

    Techniques: Cell Culture

    The effect of p21 antisense on DNA synthesis of hepatocytes in the presence of HGF. Rat hepatocytes were cultured at high density in WE containing 10% FCS, 10 ng/mL HGF, 1 mmol/L BrdU, along with various concentrations of either p21 antisense or nonsense oligonucleotide for 24 hours. An open bar denotes hepatocytes cultured in the absence of HGF. Closed bars denote hepatocytes cultured with p21 antisense oligonucleotide in the presence of 10 ng/mL HGF. A dotted bar denotes hepatocytes cultured with nonsense oligonucleotide in the presence of 10 ng/mL HGF. Data are mean ± SEM of eight dishes. *p

    Journal: PLoS ONE

    Article Title: Induction of p53-Dependent p21 Limits Proliferative Activity of Rat Hepatocytes in the Presence of Hepatocyte Growth Factor

    doi: 10.1371/journal.pone.0078346

    Figure Lengend Snippet: The effect of p21 antisense on DNA synthesis of hepatocytes in the presence of HGF. Rat hepatocytes were cultured at high density in WE containing 10% FCS, 10 ng/mL HGF, 1 mmol/L BrdU, along with various concentrations of either p21 antisense or nonsense oligonucleotide for 24 hours. An open bar denotes hepatocytes cultured in the absence of HGF. Closed bars denote hepatocytes cultured with p21 antisense oligonucleotide in the presence of 10 ng/mL HGF. A dotted bar denotes hepatocytes cultured with nonsense oligonucleotide in the presence of 10 ng/mL HGF. Data are mean ± SEM of eight dishes. *p

    Article Snippet: The suspensions were incubated with p21 antigen extraction agent (1.0 M potassium chloride, 6% zwittergent (Calbiochem, CA)), and centrifuged.

    Techniques: DNA Synthesis, Cell Culture

    Changes in hepatic p21 levels after two thirds partial hepatectomy in rats. Data are mean ± SEM of four rats. Open and closed circles indicate the hepatic p21 levels in partially hepatectomized rats and in sham-operated rats, respectively. *p

    Journal: PLoS ONE

    Article Title: Induction of p53-Dependent p21 Limits Proliferative Activity of Rat Hepatocytes in the Presence of Hepatocyte Growth Factor

    doi: 10.1371/journal.pone.0078346

    Figure Lengend Snippet: Changes in hepatic p21 levels after two thirds partial hepatectomy in rats. Data are mean ± SEM of four rats. Open and closed circles indicate the hepatic p21 levels in partially hepatectomized rats and in sham-operated rats, respectively. *p

    Article Snippet: The suspensions were incubated with p21 antigen extraction agent (1.0 M potassium chloride, 6% zwittergent (Calbiochem, CA)), and centrifuged.

    Techniques:

    Serial changes in p21 protein levels of hepatocytes treated with HGF. Rat hepatocytes were cultured at high density in WE containing 10% FCS and 10 ng/mL HGF, and were harvested serially. Closed circles denote hepatocytes cultured at high density. Open circles denote hepatocytes cultured at low density. Data are mean ± SEM of four dishes. *p

    Journal: PLoS ONE

    Article Title: Induction of p53-Dependent p21 Limits Proliferative Activity of Rat Hepatocytes in the Presence of Hepatocyte Growth Factor

    doi: 10.1371/journal.pone.0078346

    Figure Lengend Snippet: Serial changes in p21 protein levels of hepatocytes treated with HGF. Rat hepatocytes were cultured at high density in WE containing 10% FCS and 10 ng/mL HGF, and were harvested serially. Closed circles denote hepatocytes cultured at high density. Open circles denote hepatocytes cultured at low density. Data are mean ± SEM of four dishes. *p

    Article Snippet: The suspensions were incubated with p21 antigen extraction agent (1.0 M potassium chloride, 6% zwittergent (Calbiochem, CA)), and centrifuged.

    Techniques: Cell Culture

    Survival of RC rats receiving single IV 2.5 g I/kg doses of iohexol or iohexol + SBECD (sulfobutyl‐either β cyclodextrin) at mole ratio of 1:0.025 iohexol:SBECD. N = 8 in each group.

    Journal: Journal of Neuroimaging

    Article Title: Preclinical Studies of a Kidney Safe Iodinated Contrast Agent

    doi: 10.1111/jon.12356

    Figure Lengend Snippet: Survival of RC rats receiving single IV 2.5 g I/kg doses of iohexol or iohexol + SBECD (sulfobutyl‐either β cyclodextrin) at mole ratio of 1:0.025 iohexol:SBECD. N = 8 in each group.

    Article Snippet: The present report presents preclinical data for a new formulation of the most widely used iodinated contrast agent iohexol (Omnipaque from GE Healthcare) with SBECD.

    Techniques:

    Light microscopy of renal tissue of mouse (H E, PAS, 200×). Iohexol‐treated kidneys indicate pathological changes in the renal cortex (A) and medulla (C) such as tubular vacuolation, tubular dilatation (big arrow), cast formation (thin arrow), loss of brush border (arrow heads), and focal edema (E). Concurrent SBECD (sulfobutyl‐ether β cyclodextrin) administration at mole ratio 1:0.025 iohexol:SBECD significantly attenuated the morphological changes in both cortex (B) and medulla (D).

    Journal: Journal of Neuroimaging

    Article Title: Preclinical Studies of a Kidney Safe Iodinated Contrast Agent

    doi: 10.1111/jon.12356

    Figure Lengend Snippet: Light microscopy of renal tissue of mouse (H E, PAS, 200×). Iohexol‐treated kidneys indicate pathological changes in the renal cortex (A) and medulla (C) such as tubular vacuolation, tubular dilatation (big arrow), cast formation (thin arrow), loss of brush border (arrow heads), and focal edema (E). Concurrent SBECD (sulfobutyl‐ether β cyclodextrin) administration at mole ratio 1:0.025 iohexol:SBECD significantly attenuated the morphological changes in both cortex (B) and medulla (D).

    Article Snippet: The present report presents preclinical data for a new formulation of the most widely used iodinated contrast agent iohexol (Omnipaque from GE Healthcare) with SBECD.

    Techniques: Light Microscopy

    Serum creatinine levels at 24 h (mouse) or 48 h (rat) post treatment with iohexol or iohexol‐SBECD (sulfobutyl‐either β cyclodextrin) in renally compromised (RC) rodents at mole ratio 1:0.025.

    Journal: Journal of Neuroimaging

    Article Title: Preclinical Studies of a Kidney Safe Iodinated Contrast Agent

    doi: 10.1111/jon.12356

    Figure Lengend Snippet: Serum creatinine levels at 24 h (mouse) or 48 h (rat) post treatment with iohexol or iohexol‐SBECD (sulfobutyl‐either β cyclodextrin) in renally compromised (RC) rodents at mole ratio 1:0.025.

    Article Snippet: The present report presents preclinical data for a new formulation of the most widely used iodinated contrast agent iohexol (Omnipaque from GE Healthcare) with SBECD.

    Techniques:

    (A) Comparison of kidney protection by SBECD (sulfobutyl‐either β cyclodextrin) for iopamidol and iohexol in rats at mole ratio 1:0.025, iohexol:SBECD or iopamidol:SBECD. (B) Comparison of kidney protection by SBECD for iodixanol and iohexol in mice at mole ratio of 1:0.025, iohexol:SBECD or iodixanol:SBECD.

    Journal: Journal of Neuroimaging

    Article Title: Preclinical Studies of a Kidney Safe Iodinated Contrast Agent

    doi: 10.1111/jon.12356

    Figure Lengend Snippet: (A) Comparison of kidney protection by SBECD (sulfobutyl‐either β cyclodextrin) for iopamidol and iohexol in rats at mole ratio 1:0.025, iohexol:SBECD or iopamidol:SBECD. (B) Comparison of kidney protection by SBECD for iodixanol and iohexol in mice at mole ratio of 1:0.025, iohexol:SBECD or iodixanol:SBECD.

    Article Snippet: The present report presents preclinical data for a new formulation of the most widely used iodinated contrast agent iohexol (Omnipaque from GE Healthcare) with SBECD.

    Techniques: Mouse Assay

    Annexin V, Caspase‐3/7, and SYTOX Flow Cytometry . Protective effects of SBECD (sulfobutyl‐either β cyclodextrin) against iohexol‐induced apoptosis. HK‐2 cells were exposed to treatment for 1 hour with indicated more ratio of SBECD:Iohexol. Mean percentage values were calculated by averaging the results from three independent experiments ( n = 2, n = 3, and n = 3, respectively). +/‐ SEM is shown.

    Journal: Journal of Neuroimaging

    Article Title: Preclinical Studies of a Kidney Safe Iodinated Contrast Agent

    doi: 10.1111/jon.12356

    Figure Lengend Snippet: Annexin V, Caspase‐3/7, and SYTOX Flow Cytometry . Protective effects of SBECD (sulfobutyl‐either β cyclodextrin) against iohexol‐induced apoptosis. HK‐2 cells were exposed to treatment for 1 hour with indicated more ratio of SBECD:Iohexol. Mean percentage values were calculated by averaging the results from three independent experiments ( n = 2, n = 3, and n = 3, respectively). +/‐ SEM is shown.

    Article Snippet: The present report presents preclinical data for a new formulation of the most widely used iodinated contrast agent iohexol (Omnipaque from GE Healthcare) with SBECD.

    Techniques: Flow Cytometry, Cytometry

    Dose effect of SBECD (sulfobutyl‐either β cyclodextrin) addition to iohexol on mouse kidney pathology expressed as mole ratio of iohexol:SBECD. The total pathology scores were calculated by combining the six pathology parameters: tubular dilation, tubular casts, tubular vacuoles, loss of brush border, tubular degeneration, and edema.

    Journal: Journal of Neuroimaging

    Article Title: Preclinical Studies of a Kidney Safe Iodinated Contrast Agent

    doi: 10.1111/jon.12356

    Figure Lengend Snippet: Dose effect of SBECD (sulfobutyl‐either β cyclodextrin) addition to iohexol on mouse kidney pathology expressed as mole ratio of iohexol:SBECD. The total pathology scores were calculated by combining the six pathology parameters: tubular dilation, tubular casts, tubular vacuoles, loss of brush border, tubular degeneration, and edema.

    Article Snippet: The present report presents preclinical data for a new formulation of the most widely used iodinated contrast agent iohexol (Omnipaque from GE Healthcare) with SBECD.

    Techniques:

    (A) Left ventricular contractility changes following bolus dosing into the left coronary artery of iohexol or iohexol:SBECD (sulfobutyl‐either β cyclodextrin) at mole ratio of 1:0.025. (B) QTcV interval changes following bolus dosing into the left coronary artery.

    Journal: Journal of Neuroimaging

    Article Title: Preclinical Studies of a Kidney Safe Iodinated Contrast Agent

    doi: 10.1111/jon.12356

    Figure Lengend Snippet: (A) Left ventricular contractility changes following bolus dosing into the left coronary artery of iohexol or iohexol:SBECD (sulfobutyl‐either β cyclodextrin) at mole ratio of 1:0.025. (B) QTcV interval changes following bolus dosing into the left coronary artery.

    Article Snippet: The present report presents preclinical data for a new formulation of the most widely used iodinated contrast agent iohexol (Omnipaque from GE Healthcare) with SBECD.

    Techniques:

    (A) Effect of adding SBECD (sulfobutyl‐either β cyclodextrin) to iohexol on mouse outer renal cortex pathology measurements for six parameters at a mole ratio of iohexol:SBECD of 1:0.025. (B) Effect of adding SBECD to iohexol on rat outer renal cortex pathology measurements for six parameters at a mole ratio of iohexol:SBECD of 1:0.025.

    Journal: Journal of Neuroimaging

    Article Title: Preclinical Studies of a Kidney Safe Iodinated Contrast Agent

    doi: 10.1111/jon.12356

    Figure Lengend Snippet: (A) Effect of adding SBECD (sulfobutyl‐either β cyclodextrin) to iohexol on mouse outer renal cortex pathology measurements for six parameters at a mole ratio of iohexol:SBECD of 1:0.025. (B) Effect of adding SBECD to iohexol on rat outer renal cortex pathology measurements for six parameters at a mole ratio of iohexol:SBECD of 1:0.025.

    Article Snippet: The present report presents preclinical data for a new formulation of the most widely used iodinated contrast agent iohexol (Omnipaque from GE Healthcare) with SBECD.

    Techniques:

    Copine-III regulated the sensitivity of cells to sorafenib. Notes: (A) MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with indicated concentrations of sorafenib and analyzed by MTT assays. ( B ) MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with IC 50 concentration of sorafenib and analyzed by MTT assays at indicated time points. ( C ) L-02 cells, which were infected with control or CPNE3, were treated with indicated concentration of sorafenib and analyzed by MTT-assays. ( D ) MHCC97-H cells, which were infected with control or CPNE3, were treated with the IC 50 concentration of sorafenib and analyzed by MTT assays at indicated time points. * P

    Journal: Cancer Management and Research

    Article Title: Silencing the expression of copine-III enhances the sensitivity of hepatocellular carcinoma cells to the molecular targeted agent sorafenib

    doi: 10.2147/CMAR.S167781

    Figure Lengend Snippet: Copine-III regulated the sensitivity of cells to sorafenib. Notes: (A) MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with indicated concentrations of sorafenib and analyzed by MTT assays. ( B ) MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with IC 50 concentration of sorafenib and analyzed by MTT assays at indicated time points. ( C ) L-02 cells, which were infected with control or CPNE3, were treated with indicated concentration of sorafenib and analyzed by MTT-assays. ( D ) MHCC97-H cells, which were infected with control or CPNE3, were treated with the IC 50 concentration of sorafenib and analyzed by MTT assays at indicated time points. * P

    Article Snippet: The antitumor agent sorafenib (cat. no. S7397) was purchased from Selleck Company (Houston, TX, USA).

    Techniques: Infection, MTT Assay, Concentration Assay

    Silencing copine-III enhances the effect of sorafenib on MHCC97-H cell in vitro invasion and migration. Notes: MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with indicated concentrations of sorafenib and analyzed by Transwell analysis. ( A ) In vitro invasion of MHCC-97-H cells; ( B ) the in vitro migration of MHCC97-H cells. * P

    Journal: Cancer Management and Research

    Article Title: Silencing the expression of copine-III enhances the sensitivity of hepatocellular carcinoma cells to the molecular targeted agent sorafenib

    doi: 10.2147/CMAR.S167781

    Figure Lengend Snippet: Silencing copine-III enhances the effect of sorafenib on MHCC97-H cell in vitro invasion and migration. Notes: MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with indicated concentrations of sorafenib and analyzed by Transwell analysis. ( A ) In vitro invasion of MHCC-97-H cells; ( B ) the in vitro migration of MHCC97-H cells. * P

    Article Snippet: The antitumor agent sorafenib (cat. no. S7397) was purchased from Selleck Company (Houston, TX, USA).

    Techniques: In Vitro, Migration, Infection

    Endogenous mRNA level of CPNE3 in advanced hepatocellular carcinoma (HCC) tissues related to clinical outcome of patients who received sorafenib treatment. Notes: ( A and B ) Endogenous mRNA level of CPNE3 was identified by quantitative polymerase chain reaction (qPCR) as relative RNA ( A ) or represented DNA electrophoresis bands from ten representative specimens (five high and five low) ( B ). ( C ) OS of patients who received sorafenib. ( D ) Time to progress of patients who received sorafenib. ( C and D ) Survival analysis was conducted using the Kaplan–Meier and log-rank test. ( A ) * P

    Journal: Cancer Management and Research

    Article Title: Silencing the expression of copine-III enhances the sensitivity of hepatocellular carcinoma cells to the molecular targeted agent sorafenib

    doi: 10.2147/CMAR.S167781

    Figure Lengend Snippet: Endogenous mRNA level of CPNE3 in advanced hepatocellular carcinoma (HCC) tissues related to clinical outcome of patients who received sorafenib treatment. Notes: ( A and B ) Endogenous mRNA level of CPNE3 was identified by quantitative polymerase chain reaction (qPCR) as relative RNA ( A ) or represented DNA electrophoresis bands from ten representative specimens (five high and five low) ( B ). ( C ) OS of patients who received sorafenib. ( D ) Time to progress of patients who received sorafenib. ( C and D ) Survival analysis was conducted using the Kaplan–Meier and log-rank test. ( A ) * P

    Article Snippet: The antitumor agent sorafenib (cat. no. S7397) was purchased from Selleck Company (Houston, TX, USA).

    Techniques: Real-time Polymerase Chain Reaction, Nucleic Acid Electrophoresis

    Silencing copine-III enhances the effect of sorafenib on MHCC97-H cell subcutaneous growth. Notes: MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with indicated concentration of sorafenib and analyzed in a nude mice mode. The size of tumors was shown as photographs ( A ), tumor growth curve by mean ± standard deviation ( B ), or tumor weight by mean ± standard deviation ( C ). * P

    Journal: Cancer Management and Research

    Article Title: Silencing the expression of copine-III enhances the sensitivity of hepatocellular carcinoma cells to the molecular targeted agent sorafenib

    doi: 10.2147/CMAR.S167781

    Figure Lengend Snippet: Silencing copine-III enhances the effect of sorafenib on MHCC97-H cell subcutaneous growth. Notes: MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with indicated concentration of sorafenib and analyzed in a nude mice mode. The size of tumors was shown as photographs ( A ), tumor growth curve by mean ± standard deviation ( B ), or tumor weight by mean ± standard deviation ( C ). * P

    Article Snippet: The antitumor agent sorafenib (cat. no. S7397) was purchased from Selleck Company (Houston, TX, USA).

    Techniques: Infection, Concentration Assay, Mouse Assay, Standard Deviation

    Silencing copine-III enhances the effect of sorafenib on MHCC97-H cell intrahepatic growth. Notes: MHCC97-H cells, which were injected with control siRNA or siCPNE3, were injected into liver lobes to form intrahepatic nodules. Next, nude mice were treated with a solvent control or sorafenib. After 4–6 weeks of growth, tumor nodules formed by MHCC97-H cells in liver organs were examined by positron emission tomography (PET)/CT scanning. ( A ) Results showed the PET/CT images of whole animals or radioactivation of liver to blood; ( B ) images of tumor modules or PET/CT results from the liver. * P

    Journal: Cancer Management and Research

    Article Title: Silencing the expression of copine-III enhances the sensitivity of hepatocellular carcinoma cells to the molecular targeted agent sorafenib

    doi: 10.2147/CMAR.S167781

    Figure Lengend Snippet: Silencing copine-III enhances the effect of sorafenib on MHCC97-H cell intrahepatic growth. Notes: MHCC97-H cells, which were injected with control siRNA or siCPNE3, were injected into liver lobes to form intrahepatic nodules. Next, nude mice were treated with a solvent control or sorafenib. After 4–6 weeks of growth, tumor nodules formed by MHCC97-H cells in liver organs were examined by positron emission tomography (PET)/CT scanning. ( A ) Results showed the PET/CT images of whole animals or radioactivation of liver to blood; ( B ) images of tumor modules or PET/CT results from the liver. * P

    Article Snippet: The antitumor agent sorafenib (cat. no. S7397) was purchased from Selleck Company (Houston, TX, USA).

    Techniques: Injection, Mouse Assay, Positron Emission Tomography

    Silencing copine-III enhances sorafenib-induced MHCC97-H cell apoptosis. Notes: ( A – E ) MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with indicated concentrations of sorafenib and analyzed by flow cytometer. * P

    Journal: Cancer Management and Research

    Article Title: Silencing the expression of copine-III enhances the sensitivity of hepatocellular carcinoma cells to the molecular targeted agent sorafenib

    doi: 10.2147/CMAR.S167781

    Figure Lengend Snippet: Silencing copine-III enhances sorafenib-induced MHCC97-H cell apoptosis. Notes: ( A – E ) MHCC97-H cells, which were infected with control siRNA or siCPNE3, were treated with indicated concentrations of sorafenib and analyzed by flow cytometer. * P

    Article Snippet: The antitumor agent sorafenib (cat. no. S7397) was purchased from Selleck Company (Houston, TX, USA).

    Techniques: Infection, Flow Cytometry, Cytometry

    Prussian Blue staining of human chondrocytes labeled with ( A ) Endorem ® (Guerbet, Roissy, France), ( B ) dopamine-hyaluronate-maghemite nanoparticles Run IIIB/3, ( C ) hyaluronate-maghemite nanoparticles Run II/3, and ( D ) dopamine-hyaluronate-maghemite nanoparticles Run IIIC/3. Notes: Cell nuclei are counterstained with nuclear fast red. Scale bar 25 μm.

    Journal: International Journal of Nanomedicine

    Article Title: The use of dopamine-hyaluronate associate-coated maghemite nanoparticles to label cells

    doi: 10.2147/IJN.S28658

    Figure Lengend Snippet: Prussian Blue staining of human chondrocytes labeled with ( A ) Endorem ® (Guerbet, Roissy, France), ( B ) dopamine-hyaluronate-maghemite nanoparticles Run IIIB/3, ( C ) hyaluronate-maghemite nanoparticles Run II/3, and ( D ) dopamine-hyaluronate-maghemite nanoparticles Run IIIC/3. Notes: Cell nuclei are counterstained with nuclear fast red. Scale bar 25 μm.

    Article Snippet: Sodium HA (molecular weight: ~300,000) was obtained from Contipro Pharma (Dolni Dobrouc, Czech Republic), DPA hydrochloride and N -(3-dimethylaminopropyl)-N′ -ethylcarbodiimide hydrochloride from Sigma-Aldrich Corporation (St Louis, MO), and the commercial contrast agent Endorem® from Guerbet (Roissy, France).

    Techniques: Staining, Labeling

    To determine whether hyaluronate molecules, which are also present in the cartilage extracellular matrix, can affect chondrogenic differentiation, rat bone marrow mesenchymal stem cells labeled with ( A and B ) dopamine-hyaluronate-maghemite nanoparticles Run IIIC/3, ( C and D ) Endorem ® (Guerbet, Roissy, France), and ( E and F ) hyaluronate-maghemite nanoparticles Run II/3 were differentiated into a chondrogenic phenotype. Notes: The left column shows staining for Alcian Blue (a marker of chondrogenic differentiation), while the right column represents Prussian Blue staining. Iron is visible as ( A , C and E ) brown or ( B , D and F ) blue deposits.

    Journal: International Journal of Nanomedicine

    Article Title: The use of dopamine-hyaluronate associate-coated maghemite nanoparticles to label cells

    doi: 10.2147/IJN.S28658

    Figure Lengend Snippet: To determine whether hyaluronate molecules, which are also present in the cartilage extracellular matrix, can affect chondrogenic differentiation, rat bone marrow mesenchymal stem cells labeled with ( A and B ) dopamine-hyaluronate-maghemite nanoparticles Run IIIC/3, ( C and D ) Endorem ® (Guerbet, Roissy, France), and ( E and F ) hyaluronate-maghemite nanoparticles Run II/3 were differentiated into a chondrogenic phenotype. Notes: The left column shows staining for Alcian Blue (a marker of chondrogenic differentiation), while the right column represents Prussian Blue staining. Iron is visible as ( A , C and E ) brown or ( B , D and F ) blue deposits.

    Article Snippet: Sodium HA (molecular weight: ~300,000) was obtained from Contipro Pharma (Dolni Dobrouc, Czech Republic), DPA hydrochloride and N -(3-dimethylaminopropyl)-N′ -ethylcarbodiimide hydrochloride from Sigma-Aldrich Corporation (St Louis, MO), and the commercial contrast agent Endorem® from Guerbet (Roissy, France).

    Techniques: Labeling, Staining, Marker

    In the MATLAB ® 6.0 Image Processing Toolbox™ (MathWorks, Natick, MA), the color scale of Prussian Blue staining was precomputed from unstained and maximally stained nanoparticles (without cells) in CIE L*a*b color space and experimentally validated on images with stained cells. Notes: The cells in each image were manually labeled, and for each cell the staining intensity as an index on the precomputed color scale was averaged. As a result, the figure shows representative curves of the distribution of the intensity of Prussian Blue staining (X axis) of rat bone marrow mesenchymal stem cells labeled with dopamine-hyaluronate-maghemite nanoparticles Run IIIC/3, hyaluronate-maghemite nanoparticles Run II/3, and Endorem ® (Guerbet, Roissy, France). The Y axis shows the percentage of cells for each labeling intensity.

    Journal: International Journal of Nanomedicine

    Article Title: The use of dopamine-hyaluronate associate-coated maghemite nanoparticles to label cells

    doi: 10.2147/IJN.S28658

    Figure Lengend Snippet: In the MATLAB ® 6.0 Image Processing Toolbox™ (MathWorks, Natick, MA), the color scale of Prussian Blue staining was precomputed from unstained and maximally stained nanoparticles (without cells) in CIE L*a*b color space and experimentally validated on images with stained cells. Notes: The cells in each image were manually labeled, and for each cell the staining intensity as an index on the precomputed color scale was averaged. As a result, the figure shows representative curves of the distribution of the intensity of Prussian Blue staining (X axis) of rat bone marrow mesenchymal stem cells labeled with dopamine-hyaluronate-maghemite nanoparticles Run IIIC/3, hyaluronate-maghemite nanoparticles Run II/3, and Endorem ® (Guerbet, Roissy, France). The Y axis shows the percentage of cells for each labeling intensity.

    Article Snippet: Sodium HA (molecular weight: ~300,000) was obtained from Contipro Pharma (Dolni Dobrouc, Czech Republic), DPA hydrochloride and N -(3-dimethylaminopropyl)-N′ -ethylcarbodiimide hydrochloride from Sigma-Aldrich Corporation (St Louis, MO), and the commercial contrast agent Endorem® from Guerbet (Roissy, France).

    Techniques: Staining, Labeling

    Microscopic observation of Prussian Blue-stained rat bone marrow mesenchymal stem cells labeled with ( A ) dopamine-maghemite nanoparticles Run I/3, ( B ) hyaluronate-maghemite nanoparticles Run II/1 and ( C ) Run II/3, dopamine-hyaluronate-maghemite nanoparticles ( D ) Run IIIC/1, ( E ) Run IIIC/2, and ( F ) Run IIIC/3, ( G ) Endorem ® (Guerbet, Roissy, France), and ( H ) neat maghemite. Notes: Cell nuclei are counterstained with hematoxylin. Scale bar 25 μm.

    Journal: International Journal of Nanomedicine

    Article Title: The use of dopamine-hyaluronate associate-coated maghemite nanoparticles to label cells

    doi: 10.2147/IJN.S28658

    Figure Lengend Snippet: Microscopic observation of Prussian Blue-stained rat bone marrow mesenchymal stem cells labeled with ( A ) dopamine-maghemite nanoparticles Run I/3, ( B ) hyaluronate-maghemite nanoparticles Run II/1 and ( C ) Run II/3, dopamine-hyaluronate-maghemite nanoparticles ( D ) Run IIIC/1, ( E ) Run IIIC/2, and ( F ) Run IIIC/3, ( G ) Endorem ® (Guerbet, Roissy, France), and ( H ) neat maghemite. Notes: Cell nuclei are counterstained with hematoxylin. Scale bar 25 μm.

    Article Snippet: Sodium HA (molecular weight: ~300,000) was obtained from Contipro Pharma (Dolni Dobrouc, Czech Republic), DPA hydrochloride and N -(3-dimethylaminopropyl)-N′ -ethylcarbodiimide hydrochloride from Sigma-Aldrich Corporation (St Louis, MO), and the commercial contrast agent Endorem® from Guerbet (Roissy, France).

    Techniques: Staining, Labeling

    Labeling efficiency is expressed as the percentage of rat bone marrow mesenchymal stem cells labeled with hyaluronate-maghemite nanoparticles Runs II/1–3, dopamine-maghemite nanoparticles Runs I/1–3, dopamine-hyaluronate-maghemite nanoparticles Runs IIIA/1–3, IIIB/1–3, and IIIC/1–3, neat maghemite, and Endorem ® (Guerbet, Roissy, France). Notes: A colloid containing 15.4 μg of iron per mL was added to the culture media for 72 hours. All experiments were done in triplicate, counting five optical fields from each well (n = 15). Abbreviation: γ-Fe 2 O 3 , maghemite.

    Journal: International Journal of Nanomedicine

    Article Title: The use of dopamine-hyaluronate associate-coated maghemite nanoparticles to label cells

    doi: 10.2147/IJN.S28658

    Figure Lengend Snippet: Labeling efficiency is expressed as the percentage of rat bone marrow mesenchymal stem cells labeled with hyaluronate-maghemite nanoparticles Runs II/1–3, dopamine-maghemite nanoparticles Runs I/1–3, dopamine-hyaluronate-maghemite nanoparticles Runs IIIA/1–3, IIIB/1–3, and IIIC/1–3, neat maghemite, and Endorem ® (Guerbet, Roissy, France). Notes: A colloid containing 15.4 μg of iron per mL was added to the culture media for 72 hours. All experiments were done in triplicate, counting five optical fields from each well (n = 15). Abbreviation: γ-Fe 2 O 3 , maghemite.

    Article Snippet: Sodium HA (molecular weight: ~300,000) was obtained from Contipro Pharma (Dolni Dobrouc, Czech Republic), DPA hydrochloride and N -(3-dimethylaminopropyl)-N′ -ethylcarbodiimide hydrochloride from Sigma-Aldrich Corporation (St Louis, MO), and the commercial contrast agent Endorem® from Guerbet (Roissy, France).

    Techniques: Labeling

    S-nitrosylation of eNOS is increased in the penis after cavernous nerve injury, and is decreased by treatment with a denitrosylating agent NAC. eNOS S-nitrosylation was measured by TMT-switch assay consisting of immunoprecipitation of TMT-tagged proteins with anti-eNOS antibody, followed by Western blot against TMT. Upper panels are representative Western immunoblots of eNOS-SNO and β-actin in penes of Sham+Vehicle, Sham+NAC, BCNI+Vehicle, and BCNI+NAC rats (lanes 2–5). Lower panel represents quantitative analysis of eNOS-SNO/β-actin in the same treatment groups. No S-nitrosylation of eNOS could be detected in penile tissue of BCNI+Vehicle rats in the absence of ascorbate (lane 1). n=3

    Journal: International journal of impotence research

    Article Title: S-nitrosylation of NOS pathway mediators in the penis contributes to cavernous nerve injury-induced erectile dysfunction

    doi: 10.1038/s41443-018-0021-y

    Figure Lengend Snippet: S-nitrosylation of eNOS is increased in the penis after cavernous nerve injury, and is decreased by treatment with a denitrosylating agent NAC. eNOS S-nitrosylation was measured by TMT-switch assay consisting of immunoprecipitation of TMT-tagged proteins with anti-eNOS antibody, followed by Western blot against TMT. Upper panels are representative Western immunoblots of eNOS-SNO and β-actin in penes of Sham+Vehicle, Sham+NAC, BCNI+Vehicle, and BCNI+NAC rats (lanes 2–5). Lower panel represents quantitative analysis of eNOS-SNO/β-actin in the same treatment groups. No S-nitrosylation of eNOS could be detected in penile tissue of BCNI+Vehicle rats in the absence of ascorbate (lane 1). n=3

    Article Snippet: NAC, an antioxidant and a denitrosylating agent (Sigma-Aldrich, St. Louis, MO), was given to rats in drinking water starting 2 days before BCNI or sham injury and continuing for 2 weeks after the surgery.

    Techniques: Immunoprecipitation, Western Blot

    Total S-nitrosothiol and total S-nitrosylated proteins are increased in the penis after cavernous nerve injury, which is partially prevented by treatment with a denitrosylating agent NAC. Saville-Griess assay was performed without (A, for total S-nitrosothiol) or with (B, for total S-nitrosylated proteins) desalting to eliminate low molecular weight nitrosylated thiols. Each bar represents the mean ± SEM of 6 rats. *P

    Journal: International journal of impotence research

    Article Title: S-nitrosylation of NOS pathway mediators in the penis contributes to cavernous nerve injury-induced erectile dysfunction

    doi: 10.1038/s41443-018-0021-y

    Figure Lengend Snippet: Total S-nitrosothiol and total S-nitrosylated proteins are increased in the penis after cavernous nerve injury, which is partially prevented by treatment with a denitrosylating agent NAC. Saville-Griess assay was performed without (A, for total S-nitrosothiol) or with (B, for total S-nitrosylated proteins) desalting to eliminate low molecular weight nitrosylated thiols. Each bar represents the mean ± SEM of 6 rats. *P

    Article Snippet: NAC, an antioxidant and a denitrosylating agent (Sigma-Aldrich, St. Louis, MO), was given to rats in drinking water starting 2 days before BCNI or sham injury and continuing for 2 weeks after the surgery.

    Techniques: Griess Assay, Molecular Weight

    Erectile function is decreased in rats with cavernous nerve injury, which is prevented by treatment with a denitrosylating agent NAC. Sham-injured and BCNI-rats were treated with vehicle or NAC (300 mg/kg/day) starting 2 days before BCNI or sham injury and continuing for 2 weeks after the surgery. Electrical stimulation of the cavernous nerve was performed as voltage response (1, 2, and 4 V) at 16 Hz with square-wave duration of 5 msec for 1 min. Representative ICP and MAP responses to 4 V electrical stimulation (A). The stimulus interval is indicated by a solid bar. Erectile response to electrical stimulation of the cavernous nerve is indicated by maximal ICP/MAP (B) and total ICP/MAP (C). Each bar represents the mean ± SEM of 5-8 rats. *P

    Journal: International journal of impotence research

    Article Title: S-nitrosylation of NOS pathway mediators in the penis contributes to cavernous nerve injury-induced erectile dysfunction

    doi: 10.1038/s41443-018-0021-y

    Figure Lengend Snippet: Erectile function is decreased in rats with cavernous nerve injury, which is prevented by treatment with a denitrosylating agent NAC. Sham-injured and BCNI-rats were treated with vehicle or NAC (300 mg/kg/day) starting 2 days before BCNI or sham injury and continuing for 2 weeks after the surgery. Electrical stimulation of the cavernous nerve was performed as voltage response (1, 2, and 4 V) at 16 Hz with square-wave duration of 5 msec for 1 min. Representative ICP and MAP responses to 4 V electrical stimulation (A). The stimulus interval is indicated by a solid bar. Erectile response to electrical stimulation of the cavernous nerve is indicated by maximal ICP/MAP (B) and total ICP/MAP (C). Each bar represents the mean ± SEM of 5-8 rats. *P

    Article Snippet: NAC, an antioxidant and a denitrosylating agent (Sigma-Aldrich, St. Louis, MO), was given to rats in drinking water starting 2 days before BCNI or sham injury and continuing for 2 weeks after the surgery.

    Techniques:

    S-nitrosylation of sGC is increased in the penis after cavernous nerve injury, which is prevented by treatment with a denitrosylating agent NAC. sGC S-nitrosylation was measured by TMT-switch assay consisting of immunoprecipitation of TMT-tagged proteins with anti-sGC antibody, followed by Western blot against TMT. Upper panels are representative Western immunoblots of sGC-SNO and β-actin in penes of Sham+Vehicle, Sham+NAC, BCNI+Vehicle, and BCNI+NAC rats (lanes 1, 2, 4, 5). Lower panel represents quantitative analysis of sGC-SNO/β-actin in the same treatment groups. No S-nitrosylation of sGC could be detected in penile tissue of BCNI+Vehicle rats in the absence of ascorbate (lane 3). n=4.

    Journal: International journal of impotence research

    Article Title: S-nitrosylation of NOS pathway mediators in the penis contributes to cavernous nerve injury-induced erectile dysfunction

    doi: 10.1038/s41443-018-0021-y

    Figure Lengend Snippet: S-nitrosylation of sGC is increased in the penis after cavernous nerve injury, which is prevented by treatment with a denitrosylating agent NAC. sGC S-nitrosylation was measured by TMT-switch assay consisting of immunoprecipitation of TMT-tagged proteins with anti-sGC antibody, followed by Western blot against TMT. Upper panels are representative Western immunoblots of sGC-SNO and β-actin in penes of Sham+Vehicle, Sham+NAC, BCNI+Vehicle, and BCNI+NAC rats (lanes 1, 2, 4, 5). Lower panel represents quantitative analysis of sGC-SNO/β-actin in the same treatment groups. No S-nitrosylation of sGC could be detected in penile tissue of BCNI+Vehicle rats in the absence of ascorbate (lane 3). n=4.

    Article Snippet: NAC, an antioxidant and a denitrosylating agent (Sigma-Aldrich, St. Louis, MO), was given to rats in drinking water starting 2 days before BCNI or sham injury and continuing for 2 weeks after the surgery.

    Techniques: Immunoprecipitation, Western Blot

    Protein expression of P-VASP (Ser-239) is decreased in the penis after cavernous nerve injury, and is increased by treatment with a denitrosylating agent NAC. Upper panels are representative Western immunoblots of P-VASP (Ser-239)/VASP in penes of Sham+Vehicle, Sham+NAC, BCNI+Vehicle, and BCNI+NAC rats. Lower panel represents quantitative analysis of P-VASP/VASP in the same treatment groups. n=4.

    Journal: International journal of impotence research

    Article Title: S-nitrosylation of NOS pathway mediators in the penis contributes to cavernous nerve injury-induced erectile dysfunction

    doi: 10.1038/s41443-018-0021-y

    Figure Lengend Snippet: Protein expression of P-VASP (Ser-239) is decreased in the penis after cavernous nerve injury, and is increased by treatment with a denitrosylating agent NAC. Upper panels are representative Western immunoblots of P-VASP (Ser-239)/VASP in penes of Sham+Vehicle, Sham+NAC, BCNI+Vehicle, and BCNI+NAC rats. Lower panel represents quantitative analysis of P-VASP/VASP in the same treatment groups. n=4.

    Article Snippet: NAC, an antioxidant and a denitrosylating agent (Sigma-Aldrich, St. Louis, MO), was given to rats in drinking water starting 2 days before BCNI or sham injury and continuing for 2 weeks after the surgery.

    Techniques: Expressing, Western Blot

    Disrupting the interaction between MZF-1 and Elk-1 decreases EMT potential ( A ) Comparison of the gene expression profiles of upregulated (left panel) and downregulated (right panel) EMT-related genes in MZF-1 60–72 -transfected Hs578T and MB-231 cloned cells, as determined by microarray with those of the parental cells. ( B ) Immunoblotting analysis of changes in protein levels in the parental and transfected cells and in the less malignant MB-468 (MB-468) and MCF-7 cells. ( C ) Visualization and quantification of cell migration of PKCα-transfected cells by migration assay. MZF-1 60–72 construct-transfected stably cloned cells were transfected with the empty-vector or full-length PKCα construct for 3 days, and migration assay was performed. ** p

    Journal: Oncotarget

    Article Title: MZF-1/Elk-1 interaction domain as therapeutic target for protein kinase Cα-based triple-negative breast cancer cells

    doi: 10.18632/oncotarget.11337

    Figure Lengend Snippet: Disrupting the interaction between MZF-1 and Elk-1 decreases EMT potential ( A ) Comparison of the gene expression profiles of upregulated (left panel) and downregulated (right panel) EMT-related genes in MZF-1 60–72 -transfected Hs578T and MB-231 cloned cells, as determined by microarray with those of the parental cells. ( B ) Immunoblotting analysis of changes in protein levels in the parental and transfected cells and in the less malignant MB-468 (MB-468) and MCF-7 cells. ( C ) Visualization and quantification of cell migration of PKCα-transfected cells by migration assay. MZF-1 60–72 construct-transfected stably cloned cells were transfected with the empty-vector or full-length PKCα construct for 3 days, and migration assay was performed. ** p

    Article Snippet: However, although the development of therapeutic agents targeting PKCα has been the focus of several laboratories [ ], targeting PKCα often leads to off-target effects.

    Techniques: Expressing, Transfection, Clone Assay, Microarray, Migration, Construct, Stable Transfection, Plasmid Preparation

    Elk-1/MZF-1 complex binds to the promoter region of PKCα to upregulate its protein expression ( A ) Specific Elk-1/MZF-1 binding sequence at the PKCα promoter in which one of the wild-type (WT) and three of the mutated sequences were designed. Black italics denote mutated sequence. ( B ) Specific binding activities of Elk-1/MZF-1 at the PKCα promoter, as visualized by EMSA. Biotin-labeled WT probes were incubated with MB-231 cell nuclear extracts and IgG/Elk-1/MZF-1 antibodies, and the reaction was resolved on a non-denaturing polyacrylamide gel. DNA–protein complexes are denoted by black arrowheads, and the two supershifted bands are denoted by black arrows. P: probe only; V: nuclear extract only; N: nuclear extract plus probe; FP: free probe. ( C ) Verification of the specific binding activities of Elk-1/MZF-1 by a competitive assay. EMSA of biotin-labeled WT or mutated oligonucleotide probes (left panel) and competition EMSA of biotin-labeled oligonucleotides with unlabeled WT or mutated oligonucleotides in 20-fold to 100-fold molar excess (right panel) are shown. ( D ) Co-IP assay of the interaction between endogenous MZF-1 and Elk-1 in MB-231 cells. Protein extracts were IP with an anti-MZF-1 antibody or anti-Elk-1 antibody/control rabbit IgG, as indicated. The resulting immunoprecipitates were resolved by SDS-PAGE and IB with both antibodies sequentially. ( E ) Interaction between MZF-1 and Elk-1 in MB-231 cells was detected by transfection of 5 μg of either FLAG-MZF-1ΔDBD or Elk-1-c-MycΔDBD to determine the sequence requirements for protein binding. Protein extracts were IP and IB with the indicated antibodies. ( F ) Confirmation of interactions between MZF-1 and Elk-1 and DNA binding activity by ChIP and Re-ChIP assays. In the ChIP assay, chromatin was pulled down with IgG, Elk-1, and MZF-1 antibodies. In the Re-ChIP assay, the pulled-down chromatin was incubated with anti-Elk-1 antibodies, followed by anti-MZF-1 antibodies; the sequence was then reversed. The band denoted by an arrow corresponds to the amplification of the PKCα promoter by the PCR performed. The bands amplified from the total chromatin were also employed as control (Input).

    Journal: Oncotarget

    Article Title: MZF-1/Elk-1 interaction domain as therapeutic target for protein kinase Cα-based triple-negative breast cancer cells

    doi: 10.18632/oncotarget.11337

    Figure Lengend Snippet: Elk-1/MZF-1 complex binds to the promoter region of PKCα to upregulate its protein expression ( A ) Specific Elk-1/MZF-1 binding sequence at the PKCα promoter in which one of the wild-type (WT) and three of the mutated sequences were designed. Black italics denote mutated sequence. ( B ) Specific binding activities of Elk-1/MZF-1 at the PKCα promoter, as visualized by EMSA. Biotin-labeled WT probes were incubated with MB-231 cell nuclear extracts and IgG/Elk-1/MZF-1 antibodies, and the reaction was resolved on a non-denaturing polyacrylamide gel. DNA–protein complexes are denoted by black arrowheads, and the two supershifted bands are denoted by black arrows. P: probe only; V: nuclear extract only; N: nuclear extract plus probe; FP: free probe. ( C ) Verification of the specific binding activities of Elk-1/MZF-1 by a competitive assay. EMSA of biotin-labeled WT or mutated oligonucleotide probes (left panel) and competition EMSA of biotin-labeled oligonucleotides with unlabeled WT or mutated oligonucleotides in 20-fold to 100-fold molar excess (right panel) are shown. ( D ) Co-IP assay of the interaction between endogenous MZF-1 and Elk-1 in MB-231 cells. Protein extracts were IP with an anti-MZF-1 antibody or anti-Elk-1 antibody/control rabbit IgG, as indicated. The resulting immunoprecipitates were resolved by SDS-PAGE and IB with both antibodies sequentially. ( E ) Interaction between MZF-1 and Elk-1 in MB-231 cells was detected by transfection of 5 μg of either FLAG-MZF-1ΔDBD or Elk-1-c-MycΔDBD to determine the sequence requirements for protein binding. Protein extracts were IP and IB with the indicated antibodies. ( F ) Confirmation of interactions between MZF-1 and Elk-1 and DNA binding activity by ChIP and Re-ChIP assays. In the ChIP assay, chromatin was pulled down with IgG, Elk-1, and MZF-1 antibodies. In the Re-ChIP assay, the pulled-down chromatin was incubated with anti-Elk-1 antibodies, followed by anti-MZF-1 antibodies; the sequence was then reversed. The band denoted by an arrow corresponds to the amplification of the PKCα promoter by the PCR performed. The bands amplified from the total chromatin were also employed as control (Input).

    Article Snippet: However, although the development of therapeutic agents targeting PKCα has been the focus of several laboratories [ ], targeting PKCα often leads to off-target effects.

    Techniques: Expressing, Binding Assay, Sequencing, Labeling, Incubation, Co-Immunoprecipitation Assay, SDS Page, Transfection, Protein Binding, Activity Assay, Chromatin Immunoprecipitation, Amplification, Polymerase Chain Reaction

    Identification of MZF-1 and Elk-1 interacting domains ( A ) MZF-1-c-Myc and its fragments (amino acids 73–485, 1–60, 1–72, 1–141, and 60–72) were used to indicate the binding domain of MZF-1 (upper panel). Dark-black bands denote the fragments that bound Elk-1 as present in the co-immunoprecipitation assay (lower panel). HEK-293 cells were transfected with 5 μg of FLAG-Elk-1 and either empty vector or the indicated MZF-1-c-Myc fragment. The c-Myc-fused peptides were immunoprecipitated (IP) by the c-Myc monoclonal antibody and then immunoblotted (IB) with the anti-FLAG antibodies. Lysates of HEK-293 cells transfected with the FLAG-Elk-1 only were also immunoblotted as control (Input). “+” indicates the presence of a component, whereas “−“ indicates the absence of a component. ( B ) Two mutant fragments (amino acids 1–72 and 60–72) of MZF-1 in which the negatively charged aspartates in their binding domains were mutated to uncharged alanines (shown in slanted dark-black band). ( C ) Effect of normal and mutant fragments of MZF-1 on specific binding activities of Elk-1 and MZF-1 at the PKCα promoter analyzed by EMSA. The biotin-labeled WT oligonucleotide probes were incubated with a specific concentration of nuclear extracts of the different-treated HEK-293 cells. The HEK-293 cells were transfected with empty vector, Elk-1, MZF-1, MZF-1 60–72 , or mutant MZF-1 60–72 , and the reactions were resolved on a non-denaturing polyacrylamide gel. DNA–protein complexes are denoted by the black arrow-head. FP: free probe. ( D ) Elk-1-c-Myc and its fragments (amino acids 1–428, 1–86, 87–144, 87–325, 87–4281–86, 87–144, 87–325, 87–428, and 60–72) of Elk-1 used to indicate the binding domain (upper panel). Dark-black bands indicate the fragments that bound MZF-1 as present in the co-immunoprecipitation assay (lower panel). HEK-293 cells were co-transfected with 5 μg of FLAG-MZF-1 construct and either empty vector or the indicated Elk-1-c-Myc fragment. Lysates of HEK-293 cells transfected with the FLAG-MZF-1 construct were also immunoblotted as control (Input). ( E ) Two mutant Elk-1 fragments (amino acids 145–157 and 145–428) in which the negatively charged arginines in their binding domains were mutated to uncharged alanines (in slanted dark-black). ( F ) Effects of normal and mutant Elk-1 fragments on specific binding activities of Elk-1 and MZF-1 at the PKCα promoter analyzed by EMSA. Biotin-labeled wild-type oligonucleotide probes were incubated with the indicated concentration of nuclear extracts of the different-treated HEK-293 cells, which were transfected with empty vector, Elk-1, MZF-1, Elk-1 145–157 , or mutant Elk-1 145–157 .

    Journal: Oncotarget

    Article Title: MZF-1/Elk-1 interaction domain as therapeutic target for protein kinase Cα-based triple-negative breast cancer cells

    doi: 10.18632/oncotarget.11337

    Figure Lengend Snippet: Identification of MZF-1 and Elk-1 interacting domains ( A ) MZF-1-c-Myc and its fragments (amino acids 73–485, 1–60, 1–72, 1–141, and 60–72) were used to indicate the binding domain of MZF-1 (upper panel). Dark-black bands denote the fragments that bound Elk-1 as present in the co-immunoprecipitation assay (lower panel). HEK-293 cells were transfected with 5 μg of FLAG-Elk-1 and either empty vector or the indicated MZF-1-c-Myc fragment. The c-Myc-fused peptides were immunoprecipitated (IP) by the c-Myc monoclonal antibody and then immunoblotted (IB) with the anti-FLAG antibodies. Lysates of HEK-293 cells transfected with the FLAG-Elk-1 only were also immunoblotted as control (Input). “+” indicates the presence of a component, whereas “−“ indicates the absence of a component. ( B ) Two mutant fragments (amino acids 1–72 and 60–72) of MZF-1 in which the negatively charged aspartates in their binding domains were mutated to uncharged alanines (shown in slanted dark-black band). ( C ) Effect of normal and mutant fragments of MZF-1 on specific binding activities of Elk-1 and MZF-1 at the PKCα promoter analyzed by EMSA. The biotin-labeled WT oligonucleotide probes were incubated with a specific concentration of nuclear extracts of the different-treated HEK-293 cells. The HEK-293 cells were transfected with empty vector, Elk-1, MZF-1, MZF-1 60–72 , or mutant MZF-1 60–72 , and the reactions were resolved on a non-denaturing polyacrylamide gel. DNA–protein complexes are denoted by the black arrow-head. FP: free probe. ( D ) Elk-1-c-Myc and its fragments (amino acids 1–428, 1–86, 87–144, 87–325, 87–4281–86, 87–144, 87–325, 87–428, and 60–72) of Elk-1 used to indicate the binding domain (upper panel). Dark-black bands indicate the fragments that bound MZF-1 as present in the co-immunoprecipitation assay (lower panel). HEK-293 cells were co-transfected with 5 μg of FLAG-MZF-1 construct and either empty vector or the indicated Elk-1-c-Myc fragment. Lysates of HEK-293 cells transfected with the FLAG-MZF-1 construct were also immunoblotted as control (Input). ( E ) Two mutant Elk-1 fragments (amino acids 145–157 and 145–428) in which the negatively charged arginines in their binding domains were mutated to uncharged alanines (in slanted dark-black). ( F ) Effects of normal and mutant Elk-1 fragments on specific binding activities of Elk-1 and MZF-1 at the PKCα promoter analyzed by EMSA. Biotin-labeled wild-type oligonucleotide probes were incubated with the indicated concentration of nuclear extracts of the different-treated HEK-293 cells, which were transfected with empty vector, Elk-1, MZF-1, Elk-1 145–157 , or mutant Elk-1 145–157 .

    Article Snippet: However, although the development of therapeutic agents targeting PKCα has been the focus of several laboratories [ ], targeting PKCα often leads to off-target effects.

    Techniques: Binding Assay, Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation, Immunoprecipitation, Mutagenesis, Labeling, Incubation, Concentration Assay, Construct

    Disrupting the interaction between Elk-1 and MZF-1 decreases PKCα expression, drug resistance and malignant phenotypes ( A ) Changes in morphology and gene expression in MZF-1 60– 72 construct-transfected stably cloned MB-231 cells. β-actin was used as a control, and c-Myc was used as a marker for transfected cells. ( B ) Confocal microscopy showing the distribution of the Elk-1 and MZF-1 proteins. Cells were stained with antibodies against Elk-1 and MZF-1, followed by the appropriate FITC- or rhodamine-conjugated secondary antibodies. Confocal slices of 0.5 and 0.6 μm thicknesses were obtained, and images were obtained focusing the center of the nucleus. “N” denotes the nucleus; “C” denotes the cytosol. ( C ) ChIP assays indicating the binding activity of endogenous Elk-1 and MZF-1 to the PKCα promoter in various cells. The assays were performed using an MZF-1 (left panel) or Elk-1 (right panel) antibody. ( D ) Visualization and quantification of cell migration and proliferation of modified MB-231 cells. ( E ) Tumor growth in nude mice after xenografts of modified MB-231 and Hs578T cells (left panels). Tumors removed from the mice were weighed (middle images and graph) and sliced before histological examination (right images). ** p

    Journal: Oncotarget

    Article Title: MZF-1/Elk-1 interaction domain as therapeutic target for protein kinase Cα-based triple-negative breast cancer cells

    doi: 10.18632/oncotarget.11337

    Figure Lengend Snippet: Disrupting the interaction between Elk-1 and MZF-1 decreases PKCα expression, drug resistance and malignant phenotypes ( A ) Changes in morphology and gene expression in MZF-1 60– 72 construct-transfected stably cloned MB-231 cells. β-actin was used as a control, and c-Myc was used as a marker for transfected cells. ( B ) Confocal microscopy showing the distribution of the Elk-1 and MZF-1 proteins. Cells were stained with antibodies against Elk-1 and MZF-1, followed by the appropriate FITC- or rhodamine-conjugated secondary antibodies. Confocal slices of 0.5 and 0.6 μm thicknesses were obtained, and images were obtained focusing the center of the nucleus. “N” denotes the nucleus; “C” denotes the cytosol. ( C ) ChIP assays indicating the binding activity of endogenous Elk-1 and MZF-1 to the PKCα promoter in various cells. The assays were performed using an MZF-1 (left panel) or Elk-1 (right panel) antibody. ( D ) Visualization and quantification of cell migration and proliferation of modified MB-231 cells. ( E ) Tumor growth in nude mice after xenografts of modified MB-231 and Hs578T cells (left panels). Tumors removed from the mice were weighed (middle images and graph) and sliced before histological examination (right images). ** p

    Article Snippet: However, although the development of therapeutic agents targeting PKCα has been the focus of several laboratories [ ], targeting PKCα often leads to off-target effects.

    Techniques: Expressing, Construct, Transfection, Stable Transfection, Clone Assay, Marker, Confocal Microscopy, Staining, Chromatin Immunoprecipitation, Binding Assay, Activity Assay, Migration, Modification, Mouse Assay

    TAT-fused peptides decrease PKCα expression and reduce EMT ( A ) Sequences of the TAT-fused TAT-MZF-1 60–72 and TAT-Elk-1 145–157 peptides (normal and mutant). ( B ) Effects of TAT-fused peptides on cell migration. Cell migration assay was performed 3 days after peptides were added to the cells. ( C ) Immunoblotting analysis of changes in protein levels in the TAT-fused peptide-treated cells 3 days post-treatment, with β-actin as the control. ( D ) Detection of the effects of TAT-fused peptides on Elk-1 and MZF-1 interaction by co-immunoprecipitation assay. HEK-293 cells transfected with empty vector, FLAG-Elk-1, MZF-1-c-Myc, FLAG-MZF-1, or Elk-1-cMyc construct. “+” indicates the presence of each item, and “−” indicates the absence of each item. Different concentrations (50, 100, and 150 nmol) of TAT-fused peptides (normal or mutant) were then added to the lysates and incubated overnight at 4°C, and then subjected to IP with the anti-c-Myc antibody, followed by IB against FLAG and c-Myc. Lysates of HEK-293 cells transfected with FLAG-Elk-1 or FLAG-MZF-1 vector were indicated as control “Input”.

    Journal: Oncotarget

    Article Title: MZF-1/Elk-1 interaction domain as therapeutic target for protein kinase Cα-based triple-negative breast cancer cells

    doi: 10.18632/oncotarget.11337

    Figure Lengend Snippet: TAT-fused peptides decrease PKCα expression and reduce EMT ( A ) Sequences of the TAT-fused TAT-MZF-1 60–72 and TAT-Elk-1 145–157 peptides (normal and mutant). ( B ) Effects of TAT-fused peptides on cell migration. Cell migration assay was performed 3 days after peptides were added to the cells. ( C ) Immunoblotting analysis of changes in protein levels in the TAT-fused peptide-treated cells 3 days post-treatment, with β-actin as the control. ( D ) Detection of the effects of TAT-fused peptides on Elk-1 and MZF-1 interaction by co-immunoprecipitation assay. HEK-293 cells transfected with empty vector, FLAG-Elk-1, MZF-1-c-Myc, FLAG-MZF-1, or Elk-1-cMyc construct. “+” indicates the presence of each item, and “−” indicates the absence of each item. Different concentrations (50, 100, and 150 nmol) of TAT-fused peptides (normal or mutant) were then added to the lysates and incubated overnight at 4°C, and then subjected to IP with the anti-c-Myc antibody, followed by IB against FLAG and c-Myc. Lysates of HEK-293 cells transfected with FLAG-Elk-1 or FLAG-MZF-1 vector were indicated as control “Input”.

    Article Snippet: However, although the development of therapeutic agents targeting PKCα has been the focus of several laboratories [ ], targeting PKCα often leads to off-target effects.

    Techniques: Expressing, Mutagenesis, Migration, Cell Migration Assay, Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation, Construct, Incubation

    Correlations between PKCα expression and Elk-1/MZF-1 expression in breast cancer ( A ) Immunohistochemical analyses and correlations of PKCα and Elk-1/MZF-1 expression in human breast cancer. The left panel shows representative staining results for samples scored by visual assessment as “weak,” “moderate,” or “strong” according to staining intensity. The right panel depicts the numbers of each group classified based on PKCα, Elk-1, or MZF-1 staining intensity or grade. Moderate or strong expression of the genes of interest was given a positive rating, otherwise, a negative rating. Clinical characteristic grades of I, II, and III were obtained from US Biomax Inc. * P

    Journal: Oncotarget

    Article Title: MZF-1/Elk-1 interaction domain as therapeutic target for protein kinase Cα-based triple-negative breast cancer cells

    doi: 10.18632/oncotarget.11337

    Figure Lengend Snippet: Correlations between PKCα expression and Elk-1/MZF-1 expression in breast cancer ( A ) Immunohistochemical analyses and correlations of PKCα and Elk-1/MZF-1 expression in human breast cancer. The left panel shows representative staining results for samples scored by visual assessment as “weak,” “moderate,” or “strong” according to staining intensity. The right panel depicts the numbers of each group classified based on PKCα, Elk-1, or MZF-1 staining intensity or grade. Moderate or strong expression of the genes of interest was given a positive rating, otherwise, a negative rating. Clinical characteristic grades of I, II, and III were obtained from US Biomax Inc. * P

    Article Snippet: However, although the development of therapeutic agents targeting PKCα has been the focus of several laboratories [ ], targeting PKCα often leads to off-target effects.

    Techniques: Expressing, Immunohistochemistry, Staining

    A : Porcine insulin levels in the serum of C57BL mice transplanted with E42 pancreas and treated with costimulatory blockade agents (anti-LFA1, anti-CD48, and ±CTLA4-Ig), FTY720, with (◇) or without (○) debulking, at different time points after transplantation. Treatment with costimulatory antibodies was stopped at 3 months posttransplant, and graft maintenance was continued twice weekly only with FTY720. Insulin levels in the serum of NOD-SCID mice transplanted with E42 pancreas served as a positive control (♦). The inset demonstrates average pig insulin levels in transplanted mice over a course of 6 months. No statistical difference could be found between the tested groups. B : Porcine insulin levels in the serum of C57BL mice transplanted with E42 pancreas and treated with costimulatory blockade agents with or without debulking at different time points after FTY720 withdrawal. Data are presented as means ± SE.

    Journal: Diabetes

    Article Title: Pig Embryonic Pancreatic Tissue as a Source for Transplantation in Diabetes

    doi: 10.2337/db09-0112

    Figure Lengend Snippet: A : Porcine insulin levels in the serum of C57BL mice transplanted with E42 pancreas and treated with costimulatory blockade agents (anti-LFA1, anti-CD48, and ±CTLA4-Ig), FTY720, with (◇) or without (○) debulking, at different time points after transplantation. Treatment with costimulatory antibodies was stopped at 3 months posttransplant, and graft maintenance was continued twice weekly only with FTY720. Insulin levels in the serum of NOD-SCID mice transplanted with E42 pancreas served as a positive control (♦). The inset demonstrates average pig insulin levels in transplanted mice over a course of 6 months. No statistical difference could be found between the tested groups. B : Porcine insulin levels in the serum of C57BL mice transplanted with E42 pancreas and treated with costimulatory blockade agents with or without debulking at different time points after FTY720 withdrawal. Data are presented as means ± SE.

    Article Snippet: The costimulatory blockade agents (200 μg/mouse mouse CTLA4-Ig fusion protein [lot no. 20204; Chimerigen Laboratories], 250 μg/mouse anti-CD48 [hybridoma HM48-1; provided by Dr. Hideo Yagita], and anti-LFA1 [hybridoma FD441.8; Bioexpress] antibodies) were given intraperitoneally on days 0, 2, 4, and 6, and a single injection was repeated biweekly until 3 months posttransplant.

    Techniques: Mouse Assay, Transplantation Assay, Positive Control

    Porcine insulin blood levels 10 weeks after transplantation of E42 pancreas into immunosuppressed C57BL/6 mice. Recipients were treated with different combinations of costimulatory blockade agents (anti-LFA1, anti-CD48, and CTLA4-Ig) and FTY720, with or without T-cell debulking. Data are presented as means ± SE. * P ≤ 0.05.

    Journal: Diabetes

    Article Title: Pig Embryonic Pancreatic Tissue as a Source for Transplantation in Diabetes

    doi: 10.2337/db09-0112

    Figure Lengend Snippet: Porcine insulin blood levels 10 weeks after transplantation of E42 pancreas into immunosuppressed C57BL/6 mice. Recipients were treated with different combinations of costimulatory blockade agents (anti-LFA1, anti-CD48, and CTLA4-Ig) and FTY720, with or without T-cell debulking. Data are presented as means ± SE. * P ≤ 0.05.

    Article Snippet: The costimulatory blockade agents (200 μg/mouse mouse CTLA4-Ig fusion protein [lot no. 20204; Chimerigen Laboratories], 250 μg/mouse anti-CD48 [hybridoma HM48-1; provided by Dr. Hideo Yagita], and anti-LFA1 [hybridoma FD441.8; Bioexpress] antibodies) were given intraperitoneally on days 0, 2, 4, and 6, and a single injection was repeated biweekly until 3 months posttransplant.

    Techniques: Transplantation Assay, Mouse Assay

    Histological examination of E42 pig pancreatic tissue 2 weeks after transplantation under the renal capsule of C57BL with ( A ) and without ( B ) immune-suppression with costimulatory blockade (anti-LFA1, anti-CD48, and CTLA4-Ig), FTY720, and T-cell debulking. Slides were stained for pig insulin, cytokeratin (MNF116), CD3 + lymphocytes, and macrophages (F4/80), as indicated. (A high-quality representation of this figure is available in the online issue.)

    Journal: Diabetes

    Article Title: Pig Embryonic Pancreatic Tissue as a Source for Transplantation in Diabetes

    doi: 10.2337/db09-0112

    Figure Lengend Snippet: Histological examination of E42 pig pancreatic tissue 2 weeks after transplantation under the renal capsule of C57BL with ( A ) and without ( B ) immune-suppression with costimulatory blockade (anti-LFA1, anti-CD48, and CTLA4-Ig), FTY720, and T-cell debulking. Slides were stained for pig insulin, cytokeratin (MNF116), CD3 + lymphocytes, and macrophages (F4/80), as indicated. (A high-quality representation of this figure is available in the online issue.)

    Article Snippet: The costimulatory blockade agents (200 μg/mouse mouse CTLA4-Ig fusion protein [lot no. 20204; Chimerigen Laboratories], 250 μg/mouse anti-CD48 [hybridoma HM48-1; provided by Dr. Hideo Yagita], and anti-LFA1 [hybridoma FD441.8; Bioexpress] antibodies) were given intraperitoneally on days 0, 2, 4, and 6, and a single injection was repeated biweekly until 3 months posttransplant.

    Techniques: Transplantation Assay, Staining

    Histological examination of E42 pig pancreatic tissue 2–10 months after transplantation under the renal capsule of C57BL mice treated with the costimulatory blockade protocol (anti-LFA1, anti-CD48, and FTY720). Slides were stained for CD3+ lymphocytes ( A ), macrophages (F4/80) ( B ), insulin (blue) and glucagon (green) ( C ), mouse blood vessels (CD31, red), and pig blood vessels (CD31, green) ( D ), and cytokeratin (broad spectrum, red) and IgM and IgG deposits (green) ( E ). The inset in D demonstrates positive staining for pig endothelial cells (green) in E42 pancreas. The inset in E demonstrates positive normal staining for IgM and IgG deposits in the glomeruli of the kidney. The data are representative of the experiment shown in Table 1 ( n = 11). (A high-quality representation of this figure is available in the online issue.)

    Journal: Diabetes

    Article Title: Pig Embryonic Pancreatic Tissue as a Source for Transplantation in Diabetes

    doi: 10.2337/db09-0112

    Figure Lengend Snippet: Histological examination of E42 pig pancreatic tissue 2–10 months after transplantation under the renal capsule of C57BL mice treated with the costimulatory blockade protocol (anti-LFA1, anti-CD48, and FTY720). Slides were stained for CD3+ lymphocytes ( A ), macrophages (F4/80) ( B ), insulin (blue) and glucagon (green) ( C ), mouse blood vessels (CD31, red), and pig blood vessels (CD31, green) ( D ), and cytokeratin (broad spectrum, red) and IgM and IgG deposits (green) ( E ). The inset in D demonstrates positive staining for pig endothelial cells (green) in E42 pancreas. The inset in E demonstrates positive normal staining for IgM and IgG deposits in the glomeruli of the kidney. The data are representative of the experiment shown in Table 1 ( n = 11). (A high-quality representation of this figure is available in the online issue.)

    Article Snippet: The costimulatory blockade agents (200 μg/mouse mouse CTLA4-Ig fusion protein [lot no. 20204; Chimerigen Laboratories], 250 μg/mouse anti-CD48 [hybridoma HM48-1; provided by Dr. Hideo Yagita], and anti-LFA1 [hybridoma FD441.8; Bioexpress] antibodies) were given intraperitoneally on days 0, 2, 4, and 6, and a single injection was repeated biweekly until 3 months posttransplant.

    Techniques: Transplantation Assay, Mouse Assay, Staining

    Repeated systemic injections of URB597 (0.3 mg/kg daily, i.p.) reduced the development of cisplatin-evoked hyperalgesia in cisplatin-treated mice. A ) When cisplatin was co-injected with URB597, the development of mechanical hyperalgesia was delayed, and

    Journal:

    Article Title: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy

    doi: 10.1523/JNEUROSCI.0403-12.2012

    Figure Lengend Snippet: Repeated systemic injections of URB597 (0.3 mg/kg daily, i.p.) reduced the development of cisplatin-evoked hyperalgesia in cisplatin-treated mice. A ) When cisplatin was co-injected with URB597, the development of mechanical hyperalgesia was delayed, and

    Article Snippet: The chemotherapeutic agent cisplatin [ cis -diamminedichloroplatinum (II), Sigma-Aldrich, St. Louis, MO] was dissolved in sterile saline to the final concentration (1 mg/ml).

    Techniques: Mouse Assay, Injection

    Adult DRG neurons treated with cisplatin in vitro exhibited reduced neurite growth, which was prevented by co-treatment with URB597. A ) Dissociated DRG neurons cultured in the control conditions exhibited long, branched neurites with strong tubulin-ir.

    Journal:

    Article Title: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy

    doi: 10.1523/JNEUROSCI.0403-12.2012

    Figure Lengend Snippet: Adult DRG neurons treated with cisplatin in vitro exhibited reduced neurite growth, which was prevented by co-treatment with URB597. A ) Dissociated DRG neurons cultured in the control conditions exhibited long, branched neurites with strong tubulin-ir.

    Article Snippet: The chemotherapeutic agent cisplatin [ cis -diamminedichloroplatinum (II), Sigma-Aldrich, St. Louis, MO] was dissolved in sterile saline to the final concentration (1 mg/ml).

    Techniques: In Vitro, Cell Culture

    Assessment of a model of cisplatin-induced hyperalgesia

    Journal:

    Article Title: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy

    doi: 10.1523/JNEUROSCI.0403-12.2012

    Figure Lengend Snippet: Assessment of a model of cisplatin-induced hyperalgesia

    Article Snippet: The chemotherapeutic agent cisplatin [ cis -diamminedichloroplatinum (II), Sigma-Aldrich, St. Louis, MO] was dissolved in sterile saline to the final concentration (1 mg/ml).

    Techniques:

    Administration of AEA or URB597 decreased mechanical hyperalgesia produced by cisplatin. A ) A single intraplantar injection of AEA (10 μg) into a hind paw decreased withdrawal responses to a mechanical stimulus ipsilateral to the injection in

    Journal:

    Article Title: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy

    doi: 10.1523/JNEUROSCI.0403-12.2012

    Figure Lengend Snippet: Administration of AEA or URB597 decreased mechanical hyperalgesia produced by cisplatin. A ) A single intraplantar injection of AEA (10 μg) into a hind paw decreased withdrawal responses to a mechanical stimulus ipsilateral to the injection in

    Article Snippet: The chemotherapeutic agent cisplatin [ cis -diamminedichloroplatinum (II), Sigma-Aldrich, St. Louis, MO] was dissolved in sterile saline to the final concentration (1 mg/ml).

    Techniques: Produced, Injection

    URB597 attenuated effects of cisplatin in nerve. A ) Representative examples of Aα/Aβ components of the CAP from mice treated with vehicle, cisplatin, and cisplatin +URB597. Electrical stimuli are indicated by arrows and latencies of spike

    Journal:

    Article Title: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy

    doi: 10.1523/JNEUROSCI.0403-12.2012

    Figure Lengend Snippet: URB597 attenuated effects of cisplatin in nerve. A ) Representative examples of Aα/Aβ components of the CAP from mice treated with vehicle, cisplatin, and cisplatin +URB597. Electrical stimuli are indicated by arrows and latencies of spike

    Article Snippet: The chemotherapeutic agent cisplatin [ cis -diamminedichloroplatinum (II), Sigma-Aldrich, St. Louis, MO] was dissolved in sterile saline to the final concentration (1 mg/ml).

    Techniques: Mouse Assay

    URB597 attenuated effects of cisplatin on protein expression in DRGs. A ) TRPV1-and ATF-3-ir were detected by immunofluorescence in L3–L5 DRGs from mice treated with vehicle, cisplatin or cisplatin+URB597. Cisplatin (1 mg/kg of body weight, daily

    Journal:

    Article Title: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy

    doi: 10.1523/JNEUROSCI.0403-12.2012

    Figure Lengend Snippet: URB597 attenuated effects of cisplatin on protein expression in DRGs. A ) TRPV1-and ATF-3-ir were detected by immunofluorescence in L3–L5 DRGs from mice treated with vehicle, cisplatin or cisplatin+URB597. Cisplatin (1 mg/kg of body weight, daily

    Article Snippet: The chemotherapeutic agent cisplatin [ cis -diamminedichloroplatinum (II), Sigma-Aldrich, St. Louis, MO] was dissolved in sterile saline to the final concentration (1 mg/ml).

    Techniques: Expressing, Immunofluorescence, Mouse Assay

    Cytotoxicity of cisplatin in vitro was confirmed on cultured fibrosarcoma cells. Treatment with cisplatin (4 μg/ml for 24 h) decreased the number of viable fibrosarcoma cells. Cultures were incubated in acridine orange (0.67 μM for 5 min)

    Journal:

    Article Title: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy

    doi: 10.1523/JNEUROSCI.0403-12.2012

    Figure Lengend Snippet: Cytotoxicity of cisplatin in vitro was confirmed on cultured fibrosarcoma cells. Treatment with cisplatin (4 μg/ml for 24 h) decreased the number of viable fibrosarcoma cells. Cultures were incubated in acridine orange (0.67 μM for 5 min)

    Article Snippet: The chemotherapeutic agent cisplatin [ cis -diamminedichloroplatinum (II), Sigma-Aldrich, St. Louis, MO] was dissolved in sterile saline to the final concentration (1 mg/ml).

    Techniques: In Vitro, Cell Culture, Incubation

    URB597 attenuated a neurotoxic effect of cisplatin on DRG neurons in vitro

    Journal:

    Article Title: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy

    doi: 10.1523/JNEUROSCI.0403-12.2012

    Figure Lengend Snippet: URB597 attenuated a neurotoxic effect of cisplatin on DRG neurons in vitro

    Article Snippet: The chemotherapeutic agent cisplatin [ cis -diamminedichloroplatinum (II), Sigma-Aldrich, St. Louis, MO] was dissolved in sterile saline to the final concentration (1 mg/ml).

    Techniques: In Vitro

    Assessment of a model of cisplatin-induced mechanical hyperalgesia. A ) Weight loss was observed only in mice given the highest daily dose of cisplatin (3 mg/kg). At doses of 0.5 mg/kg or 1 mg/kg cisplatin did not produce weight loss (B-baseline). B ) Mechanical

    Journal:

    Article Title: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy

    doi: 10.1523/JNEUROSCI.0403-12.2012

    Figure Lengend Snippet: Assessment of a model of cisplatin-induced mechanical hyperalgesia. A ) Weight loss was observed only in mice given the highest daily dose of cisplatin (3 mg/kg). At doses of 0.5 mg/kg or 1 mg/kg cisplatin did not produce weight loss (B-baseline). B ) Mechanical

    Article Snippet: The chemotherapeutic agent cisplatin [ cis -diamminedichloroplatinum (II), Sigma-Aldrich, St. Louis, MO] was dissolved in sterile saline to the final concentration (1 mg/ml).

    Techniques: Mouse Assay

    Dual energy micro-CT material decomposition. (A) , In vitro phantom consisting of a large tube of water surrounded by vials containing gold, iodine, or a mixture of the two. (B) , In vivo imaging of gold nanoparticles and iodine-containing liposomes within a mouse soft tissue sarcoma. The iodine (shown in red) and gold (shown in green) maps are the result of dual energy decomposition. In both cases, the decomposition was able to successfully differentiate the signals from the gold and iodine contrast agents.

    Journal: Frontiers in Pharmacology

    Article Title: In vivo small animal micro-CT using nanoparticle contrast agents

    doi: 10.3389/fphar.2015.00256

    Figure Lengend Snippet: Dual energy micro-CT material decomposition. (A) , In vitro phantom consisting of a large tube of water surrounded by vials containing gold, iodine, or a mixture of the two. (B) , In vivo imaging of gold nanoparticles and iodine-containing liposomes within a mouse soft tissue sarcoma. The iodine (shown in red) and gold (shown in green) maps are the result of dual energy decomposition. In both cases, the decomposition was able to successfully differentiate the signals from the gold and iodine contrast agents.

    Article Snippet: Some metal nanoparticle contrast agents are commercially available, including the gold nanoparticle agent AuroVistTM (Nanoprobes, Inc., Yaphank, NY, USA) and the barium nanoparticle agent ExitronTM Nano (Miltenyi Biotec).

    Techniques: Micro-CT, In Vitro, In Vivo Imaging

    Longitudinal micro-CT imaging of liver metastases in a mouse following injection of a nanoparticle contrast agent. A single mouse is shown at 9 days (A) , 12 days (B) , 14 days (C) , and 19 days (D) after intrasplenic injection of tumor cells. Normal liver tissue is highly enhancing due to nanoparticle uptake, while the tumor regions show no enhancement. The enhancement remains high within the normal liver over the entire course of the experiment. By day 19, metastatic tumors take up the majority of the liver volume. Reprinted from ( Boll et al., 2011 ) under the Creative Commons Attribution License.

    Journal: Frontiers in Pharmacology

    Article Title: In vivo small animal micro-CT using nanoparticle contrast agents

    doi: 10.3389/fphar.2015.00256

    Figure Lengend Snippet: Longitudinal micro-CT imaging of liver metastases in a mouse following injection of a nanoparticle contrast agent. A single mouse is shown at 9 days (A) , 12 days (B) , 14 days (C) , and 19 days (D) after intrasplenic injection of tumor cells. Normal liver tissue is highly enhancing due to nanoparticle uptake, while the tumor regions show no enhancement. The enhancement remains high within the normal liver over the entire course of the experiment. By day 19, metastatic tumors take up the majority of the liver volume. Reprinted from ( Boll et al., 2011 ) under the Creative Commons Attribution License.

    Article Snippet: Some metal nanoparticle contrast agents are commercially available, including the gold nanoparticle agent AuroVistTM (Nanoprobes, Inc., Yaphank, NY, USA) and the barium nanoparticle agent ExitronTM Nano (Miltenyi Biotec).

    Techniques: Micro-CT, Imaging, Injection

    Spectral micro-CT imaging using photon counting x-ray detectors and HDL-encapsulated gold nanoparticles. (A) In vitro aorta phantom study demonstrating the conventional CT image along with the decomposition of the CT image into gold, iodine, photoelectric, and Compton components. (B) In vivo imaging of targeted gold nanoparticles and blood pool iodine in a mouse model of atherosclerosis. The iodine (red) can be clearly visualized within the aorta, while the gold signal (yellow) is immediately adjacent to the aorta lumen in the atherosclerotic plaque. Reprinted with permission from ( Cormode et al., 2010 ).

    Journal: Frontiers in Pharmacology

    Article Title: In vivo small animal micro-CT using nanoparticle contrast agents

    doi: 10.3389/fphar.2015.00256

    Figure Lengend Snippet: Spectral micro-CT imaging using photon counting x-ray detectors and HDL-encapsulated gold nanoparticles. (A) In vitro aorta phantom study demonstrating the conventional CT image along with the decomposition of the CT image into gold, iodine, photoelectric, and Compton components. (B) In vivo imaging of targeted gold nanoparticles and blood pool iodine in a mouse model of atherosclerosis. The iodine (red) can be clearly visualized within the aorta, while the gold signal (yellow) is immediately adjacent to the aorta lumen in the atherosclerotic plaque. Reprinted with permission from ( Cormode et al., 2010 ).

    Article Snippet: Some metal nanoparticle contrast agents are commercially available, including the gold nanoparticle agent AuroVistTM (Nanoprobes, Inc., Yaphank, NY, USA) and the barium nanoparticle agent ExitronTM Nano (Miltenyi Biotec).

    Techniques: Micro-CT, Imaging, In Vitro, In Vivo Imaging

    Iodine-containing nanoparticles. (A) Representation of individual amphiphilic lipids that can be incorporated into nanoparticles. (B) Representation of several configurations of self-assembling nanoparticles based on amphiphilic lipids. Iodine can be incorporated into the hydrophobic portion of the micelle, within the non-polar core of an emulsion, or within the aqueous core of liposomes. Reprinted with permission from ( Mulder et al., 2006 ).

    Journal: Frontiers in Pharmacology

    Article Title: In vivo small animal micro-CT using nanoparticle contrast agents

    doi: 10.3389/fphar.2015.00256

    Figure Lengend Snippet: Iodine-containing nanoparticles. (A) Representation of individual amphiphilic lipids that can be incorporated into nanoparticles. (B) Representation of several configurations of self-assembling nanoparticles based on amphiphilic lipids. Iodine can be incorporated into the hydrophobic portion of the micelle, within the non-polar core of an emulsion, or within the aqueous core of liposomes. Reprinted with permission from ( Mulder et al., 2006 ).

    Article Snippet: Some metal nanoparticle contrast agents are commercially available, including the gold nanoparticle agent AuroVistTM (Nanoprobes, Inc., Yaphank, NY, USA) and the barium nanoparticle agent ExitronTM Nano (Miltenyi Biotec).

    Techniques:

    3D micro-CT reconstructions of mice with EGFR-expressing tumors. Mice were injected with (A) saline, (B) non-targeted gold nanoparticles, or (C) EGFR-antibody targeted gold nanoparticles. Increased CT enhancement was seen for both types of nanoparticles, but targeted nanoparticles showed significantly higher enhancement than non-targeted controls. Reprinted from with permission from ( Reuveni et al., 2011a )

    Journal: Frontiers in Pharmacology

    Article Title: In vivo small animal micro-CT using nanoparticle contrast agents

    doi: 10.3389/fphar.2015.00256

    Figure Lengend Snippet: 3D micro-CT reconstructions of mice with EGFR-expressing tumors. Mice were injected with (A) saline, (B) non-targeted gold nanoparticles, or (C) EGFR-antibody targeted gold nanoparticles. Increased CT enhancement was seen for both types of nanoparticles, but targeted nanoparticles showed significantly higher enhancement than non-targeted controls. Reprinted from with permission from ( Reuveni et al., 2011a )

    Article Snippet: Some metal nanoparticle contrast agents are commercially available, including the gold nanoparticle agent AuroVistTM (Nanoprobes, Inc., Yaphank, NY, USA) and the barium nanoparticle agent ExitronTM Nano (Miltenyi Biotec).

    Techniques: Micro-CT, Mouse Assay, Expressing, Injection

    Three-energy micro-CT imaging in a mouse. Liposomal iodine was injected 72 h before imaging. Gold nanoparticles and low molecular weight gadolinium were injected immediately before imaging. Images were acquired at three energies, filtered, then separated into maps of iodine (red), gold (green), and gadolinium (blue) concentration.

    Journal: Frontiers in Pharmacology

    Article Title: In vivo small animal micro-CT using nanoparticle contrast agents

    doi: 10.3389/fphar.2015.00256

    Figure Lengend Snippet: Three-energy micro-CT imaging in a mouse. Liposomal iodine was injected 72 h before imaging. Gold nanoparticles and low molecular weight gadolinium were injected immediately before imaging. Images were acquired at three energies, filtered, then separated into maps of iodine (red), gold (green), and gadolinium (blue) concentration.

    Article Snippet: Some metal nanoparticle contrast agents are commercially available, including the gold nanoparticle agent AuroVistTM (Nanoprobes, Inc., Yaphank, NY, USA) and the barium nanoparticle agent ExitronTM Nano (Miltenyi Biotec).

    Techniques: Micro-CT, Imaging, Injection, Molecular Weight, Concentration Assay

    Dual blockade of HER3 and EGFR can effectively inhibit the proliferation of Ctx R clones. (A) Combinatorial treatment of Ctx R clones with cetuximab and U3-1287 leads to proliferation inhibition. Cell proliferation was measured using crystal violet assay and plotted as a percentage of proliferation relative to the vehicle control cells. Data points are represented as mean ± s.e.m. (n = 3). (B) Combinatorial treatment with cetuximab and U3-1287 leads to loss of HER3 expression in Ctx R clones. Protein lysates were fractionated on SDS–PAGE followed by immunoblotting for the indicated proteins. α-Tubulin was used as a loading control.

    Journal: Molecular Cancer

    Article Title: Overcoming acquired resistance to cetuximab by dual targeting HER family receptors with antibody-based therapy

    doi: 10.1186/1476-4598-13-242

    Figure Lengend Snippet: Dual blockade of HER3 and EGFR can effectively inhibit the proliferation of Ctx R clones. (A) Combinatorial treatment of Ctx R clones with cetuximab and U3-1287 leads to proliferation inhibition. Cell proliferation was measured using crystal violet assay and plotted as a percentage of proliferation relative to the vehicle control cells. Data points are represented as mean ± s.e.m. (n = 3). (B) Combinatorial treatment with cetuximab and U3-1287 leads to loss of HER3 expression in Ctx R clones. Protein lysates were fractionated on SDS–PAGE followed by immunoblotting for the indicated proteins. α-Tubulin was used as a loading control.

    Article Snippet: CtxR cell line (HC4) was analyzed in the panel of phosphorylation profiles of kinases after treatment with cetuximab, U3-1287 and a combination of cetuximab and U3-1287 agents (Human Phospho-Kinase Array, ARY003; R & D Systems).

    Techniques: Clone Assay, Inhibition, Crystal Violet Assay, Expressing, SDS Page

    Combination of cetuximab and U3-1287 treatment of Ctx R tumors leads to growth delay in vivo. (A) Growth-delay effects of U3-1287 in Ctx R tumors in vivo. The black arrow designates the starting time point of U3-1287 treatment. The average tumor volume of mice treated with IgG is included in all groups for comparison purposes. (B) Combination treatment with cetuximab and U3-1287 inhibited HER3 expression and HER2 activation in vivo. Total and phosphorylation levels of HER2 and HER3 proteins in Ctx R xenograft tumors were determined by immunoblot analysis after cetuximab, U3-1287 or the combination of cetuximab and U3-1287 treatments. (C) The inhibition of phospho-HER3 and phospho-HER2 expression in Ctx R tumors after combinatorial treatment corresponds with reduced proliferation and increased apoptosis. Ctx R tumor samples after cetuximab, U3-1287 or the combination of cetuximab and U3-1287 treatment in vivo were prepared and analyzed for Ki67 and cleaved caspase 3 by immunohistochemistry. Images were quantified via taking the average staining intensity measured from 3 tumors per treatment group (3 images/tumor, n = 9). Magnification 100X.

    Journal: Molecular Cancer

    Article Title: Overcoming acquired resistance to cetuximab by dual targeting HER family receptors with antibody-based therapy

    doi: 10.1186/1476-4598-13-242

    Figure Lengend Snippet: Combination of cetuximab and U3-1287 treatment of Ctx R tumors leads to growth delay in vivo. (A) Growth-delay effects of U3-1287 in Ctx R tumors in vivo. The black arrow designates the starting time point of U3-1287 treatment. The average tumor volume of mice treated with IgG is included in all groups for comparison purposes. (B) Combination treatment with cetuximab and U3-1287 inhibited HER3 expression and HER2 activation in vivo. Total and phosphorylation levels of HER2 and HER3 proteins in Ctx R xenograft tumors were determined by immunoblot analysis after cetuximab, U3-1287 or the combination of cetuximab and U3-1287 treatments. (C) The inhibition of phospho-HER3 and phospho-HER2 expression in Ctx R tumors after combinatorial treatment corresponds with reduced proliferation and increased apoptosis. Ctx R tumor samples after cetuximab, U3-1287 or the combination of cetuximab and U3-1287 treatment in vivo were prepared and analyzed for Ki67 and cleaved caspase 3 by immunohistochemistry. Images were quantified via taking the average staining intensity measured from 3 tumors per treatment group (3 images/tumor, n = 9). Magnification 100X.

    Article Snippet: CtxR cell line (HC4) was analyzed in the panel of phosphorylation profiles of kinases after treatment with cetuximab, U3-1287 and a combination of cetuximab and U3-1287 agents (Human Phospho-Kinase Array, ARY003; R & D Systems).

    Techniques: In Vivo, Mouse Assay, Expressing, Activation Assay, Inhibition, Immunohistochemistry, Staining

    U3-1287 downregulates total and phosphorylation of HER3 as well as AKT phosphorylation, but does not inhibit cell proliferation in Ctx R clones. (A) U3-1287 alone did not inhibit cell proliferation in Ctx R clones. The cell proliferation was measured via crystal violet assay and plotted as a percentage of proliferation relative to the vehicle control cells. Data points are represented as mean ± s.e.m. (n = 3). (B) U3-1287 downregulates total HER3 and phosphorylation of AKT in Ctx R clones. Whole cell lysates were fractionated on SDS-PAGE followed by immunoblotting for the indicated proteins. α-Tubulin was used as a loading control.

    Journal: Molecular Cancer

    Article Title: Overcoming acquired resistance to cetuximab by dual targeting HER family receptors with antibody-based therapy

    doi: 10.1186/1476-4598-13-242

    Figure Lengend Snippet: U3-1287 downregulates total and phosphorylation of HER3 as well as AKT phosphorylation, but does not inhibit cell proliferation in Ctx R clones. (A) U3-1287 alone did not inhibit cell proliferation in Ctx R clones. The cell proliferation was measured via crystal violet assay and plotted as a percentage of proliferation relative to the vehicle control cells. Data points are represented as mean ± s.e.m. (n = 3). (B) U3-1287 downregulates total HER3 and phosphorylation of AKT in Ctx R clones. Whole cell lysates were fractionated on SDS-PAGE followed by immunoblotting for the indicated proteins. α-Tubulin was used as a loading control.

    Article Snippet: CtxR cell line (HC4) was analyzed in the panel of phosphorylation profiles of kinases after treatment with cetuximab, U3-1287 and a combination of cetuximab and U3-1287 agents (Human Phospho-Kinase Array, ARY003; R & D Systems).

    Techniques: Clone Assay, Crystal Violet Assay, SDS Page

    Combined treatment of Ctx R clones with cetuximab and U3-1287 inhibits HER2, AKT and MAPK signalings more effectively than either drug alone. (A) Human Phospho-Kinase array analysis demonstrated that combined treatment with cetuximab and U3-1287 inhibits proliferation and survival signaling in Ctx R cell clone, HC4. The cell extracts were incubated with membranes containing antibodies to 46 different kinase phosphorylation sites. Quantitation of phosphorylated proteins was completed using scanned images from ImageJ software. Data points are represented as the mean of duplicate spots. (B) Effects of combined cetuximab and U3-1287 treatment on their respective kinase targets in Ctx R clones. Protein lysates from Figure 5A (HC4) were fractionated on SDS–PAGE followed by immunoblotting for the indicated proteins. Protein lysate from other Ctx R clones (HC1 and HC8) as well as HP cells were obtained after treatment with vehicle, cetuximab (20 ug/mL), U3-1287 (100 ug/mL) or the combination of cetuximab and U3-1287 for 24 h. α-Tubulin was used as a loading control.

    Journal: Molecular Cancer

    Article Title: Overcoming acquired resistance to cetuximab by dual targeting HER family receptors with antibody-based therapy

    doi: 10.1186/1476-4598-13-242

    Figure Lengend Snippet: Combined treatment of Ctx R clones with cetuximab and U3-1287 inhibits HER2, AKT and MAPK signalings more effectively than either drug alone. (A) Human Phospho-Kinase array analysis demonstrated that combined treatment with cetuximab and U3-1287 inhibits proliferation and survival signaling in Ctx R cell clone, HC4. The cell extracts were incubated with membranes containing antibodies to 46 different kinase phosphorylation sites. Quantitation of phosphorylated proteins was completed using scanned images from ImageJ software. Data points are represented as the mean of duplicate spots. (B) Effects of combined cetuximab and U3-1287 treatment on their respective kinase targets in Ctx R clones. Protein lysates from Figure 5A (HC4) were fractionated on SDS–PAGE followed by immunoblotting for the indicated proteins. Protein lysate from other Ctx R clones (HC1 and HC8) as well as HP cells were obtained after treatment with vehicle, cetuximab (20 ug/mL), U3-1287 (100 ug/mL) or the combination of cetuximab and U3-1287 for 24 h. α-Tubulin was used as a loading control.

    Article Snippet: CtxR cell line (HC4) was analyzed in the panel of phosphorylation profiles of kinases after treatment with cetuximab, U3-1287 and a combination of cetuximab and U3-1287 agents (Human Phospho-Kinase Array, ARY003; R & D Systems).

    Techniques: Clone Assay, Incubation, Quantitation Assay, Software, SDS Page

    Cetuximab and U3-1287 induced apoptosis in Ctx R clones. (A) Combinatorial treatment with cetuximab and U3-1287 activates Caspase 3/7 compared to either drug alone in Ctx R clones. Caspase-3/7 activity was determined by Caspase 3/7-Glo assay. Data represent means ± s.e.m from 3 independent experiments (n = 9). *p

    Journal: Molecular Cancer

    Article Title: Overcoming acquired resistance to cetuximab by dual targeting HER family receptors with antibody-based therapy

    doi: 10.1186/1476-4598-13-242

    Figure Lengend Snippet: Cetuximab and U3-1287 induced apoptosis in Ctx R clones. (A) Combinatorial treatment with cetuximab and U3-1287 activates Caspase 3/7 compared to either drug alone in Ctx R clones. Caspase-3/7 activity was determined by Caspase 3/7-Glo assay. Data represent means ± s.e.m from 3 independent experiments (n = 9). *p

    Article Snippet: CtxR cell line (HC4) was analyzed in the panel of phosphorylation profiles of kinases after treatment with cetuximab, U3-1287 and a combination of cetuximab and U3-1287 agents (Human Phospho-Kinase Array, ARY003; R & D Systems).

    Techniques: Clone Assay, Activity Assay, Glo Assay

    Elevated IOP increases TRPV1 in DBA/2 mice. Immunolabeling against TRPV1 with DAPI counterstain in retina from DBA/2 mice ( A, B ) and colabeling of the RGC-specific marker Thy1 and TRPV1 C57 retina for comparison ( C–E ). All micrographs from the

    Journal:

    Article Title: TRPV1: Contribution to Retinal Ganglion Cell Apoptosis and Increased Intracellular Ca2+ with Exposure to Hydrostatic Pressure

    doi: 10.1167/iovs.08-2321

    Figure Lengend Snippet: Elevated IOP increases TRPV1 in DBA/2 mice. Immunolabeling against TRPV1 with DAPI counterstain in retina from DBA/2 mice ( A, B ) and colabeling of the RGC-specific marker Thy1 and TRPV1 C57 retina for comparison ( C–E ). All micrographs from the

    Article Snippet: – For TRPV1-specific agonism, we again used a widely accepted TRPV1-specific agent capsaicin (Sigma).

    Techniques: Mouse Assay, Immunolabeling, Marker

    TRPV1 localization in RGCs of rat retina. ( A ) Immunocytochemical labeling for TRPV1 shows strong localization in the outer retina and in RGCs (large cell bodies); there is little or no label in smaller displaced amacrine cells of the GCL. Clear examples

    Journal:

    Article Title: TRPV1: Contribution to Retinal Ganglion Cell Apoptosis and Increased Intracellular Ca2+ with Exposure to Hydrostatic Pressure

    doi: 10.1167/iovs.08-2321

    Figure Lengend Snippet: TRPV1 localization in RGCs of rat retina. ( A ) Immunocytochemical labeling for TRPV1 shows strong localization in the outer retina and in RGCs (large cell bodies); there is little or no label in smaller displaced amacrine cells of the GCL. Clear examples

    Article Snippet: – For TRPV1-specific agonism, we again used a widely accepted TRPV1-specific agent capsaicin (Sigma).

    Techniques: Labeling

    Antagonism of TRPV1 diminishes pressure-induced RGC apoptosis. ( A ) Density and ( B ) percentage of TUNEL-positive RGCs at ambient or elevated pressure for 48 hours exposed to increasing concentrations of I-RTX. ( A ) For vehicle only, elevated pressure reduces

    Journal:

    Article Title: TRPV1: Contribution to Retinal Ganglion Cell Apoptosis and Increased Intracellular Ca2+ with Exposure to Hydrostatic Pressure

    doi: 10.1167/iovs.08-2321

    Figure Lengend Snippet: Antagonism of TRPV1 diminishes pressure-induced RGC apoptosis. ( A ) Density and ( B ) percentage of TUNEL-positive RGCs at ambient or elevated pressure for 48 hours exposed to increasing concentrations of I-RTX. ( A ) For vehicle only, elevated pressure reduces

    Article Snippet: – For TRPV1-specific agonism, we again used a widely accepted TRPV1-specific agent capsaicin (Sigma).

    Techniques: TUNEL Assay

    Activation of TRPV1 induces RGC apoptosis. Density and percentage of TUNEL-positive RGCs in primary cultures exposed to varying concentrations of the TRPV1 agonist capsaicin (1–100 µM) for 48 hours. As capsaicin concentration increases,

    Journal:

    Article Title: TRPV1: Contribution to Retinal Ganglion Cell Apoptosis and Increased Intracellular Ca2+ with Exposure to Hydrostatic Pressure

    doi: 10.1167/iovs.08-2321

    Figure Lengend Snippet: Activation of TRPV1 induces RGC apoptosis. Density and percentage of TUNEL-positive RGCs in primary cultures exposed to varying concentrations of the TRPV1 agonist capsaicin (1–100 µM) for 48 hours. As capsaicin concentration increases,

    Article Snippet: – For TRPV1-specific agonism, we again used a widely accepted TRPV1-specific agent capsaicin (Sigma).

    Techniques: Activation Assay, TUNEL Assay, Concentration Assay

    TRPV1 contributes to pressure-induced increases in RGC Ca 2+ . ( A ) Fluo-4 conjugated Ca 2+ accumulation in RGCs at ambient pressure ( left ) and quantification with intensity map ( right ). ( B ) One hour of elevated hydrostatic pressure induces increased intracellular

    Journal:

    Article Title: TRPV1: Contribution to Retinal Ganglion Cell Apoptosis and Increased Intracellular Ca2+ with Exposure to Hydrostatic Pressure

    doi: 10.1167/iovs.08-2321

    Figure Lengend Snippet: TRPV1 contributes to pressure-induced increases in RGC Ca 2+ . ( A ) Fluo-4 conjugated Ca 2+ accumulation in RGCs at ambient pressure ( left ) and quantification with intensity map ( right ). ( B ) One hour of elevated hydrostatic pressure induces increased intracellular

    Article Snippet: – For TRPV1-specific agonism, we again used a widely accepted TRPV1-specific agent capsaicin (Sigma).

    Techniques:

    Expression of trpv1 mRNA in RGCs. ( A ) PCR demonstrates expression of trpV1 mRNA in whole retina from adult rat, RGCs isolated from adult and P4 rat retina, and P3 RGCs cultured for 7 days. ( B ) Control in situ hybridization with sense trpv1 probe reveals

    Journal:

    Article Title: TRPV1: Contribution to Retinal Ganglion Cell Apoptosis and Increased Intracellular Ca2+ with Exposure to Hydrostatic Pressure

    doi: 10.1167/iovs.08-2321

    Figure Lengend Snippet: Expression of trpv1 mRNA in RGCs. ( A ) PCR demonstrates expression of trpV1 mRNA in whole retina from adult rat, RGCs isolated from adult and P4 rat retina, and P3 RGCs cultured for 7 days. ( B ) Control in situ hybridization with sense trpv1 probe reveals

    Article Snippet: – For TRPV1-specific agonism, we again used a widely accepted TRPV1-specific agent capsaicin (Sigma).

    Techniques: Expressing, Polymerase Chain Reaction, Isolation, Cell Culture, In Situ Hybridization

    Identification of tivantinib and ABT-199 as a synergistic drug combination in AML cells. ( a ) Results of tivantinib combination drug screen in HL60 cells using a customized library of 240 targeted agents. Replicate correlations of cell viability following treatment with individual library compounds (2.5 μM) (left) and compounds in combination with tivantinib (0.25 μM) (middle) are displayed. Fold change corresponds to the ratio of inhibition of cell viability achieved by a drug combination with tivantinib (0.25 μM) compared to individual single library compounds (2.5 μM). Drugs passing fold change > 1.5 cutoff are highlighted in red. Navitoclax and ABT-199 are labeled. ( b ) Dose response curves for inhibition of viability of HL60 cells of tivantinib and its combination with either ABT-199 (left) or cytarabine (Ara-C; right). Synergy is assessed by the Bliss model of independence (histograms in insets). Displayed in the histograms are the experimentally determined differences for each drug combination from the calculated Bliss additivity on a scale of +20% to −20% cell viability in order of increasing tivantinib concentrations. Vertical lines indicate increments of 10% cell viability. Bars pointing up from the blue baseline (additivity) indicate synergy, bars pointing down indicate antagonism. ( c ) Effects of tivantinib and ABT-199 combination (in μM) on PARP-1 and caspase 3 cleavage as well as pSer10 histone H3 levels after 24 h treatment. ( d ) Effects of tivantinib and ABT-199 combination on β-catenin stabilization and pGSK3α/β Y279/216 levels. ( e ) Effects of tivantinib and ABT-199 combination on MCL-1, BCL-XL, and Bak. V = vehicle (DMSO). Tivantinib, ABT-199, and BIO concentrations are in μM. NaCl and LiCl concentrations are in mM.

    Journal: Scientific Reports

    Article Title: Off-target based drug repurposing opportunities for tivantinib in acute myeloid leukemia

    doi: 10.1038/s41598-018-37174-6

    Figure Lengend Snippet: Identification of tivantinib and ABT-199 as a synergistic drug combination in AML cells. ( a ) Results of tivantinib combination drug screen in HL60 cells using a customized library of 240 targeted agents. Replicate correlations of cell viability following treatment with individual library compounds (2.5 μM) (left) and compounds in combination with tivantinib (0.25 μM) (middle) are displayed. Fold change corresponds to the ratio of inhibition of cell viability achieved by a drug combination with tivantinib (0.25 μM) compared to individual single library compounds (2.5 μM). Drugs passing fold change > 1.5 cutoff are highlighted in red. Navitoclax and ABT-199 are labeled. ( b ) Dose response curves for inhibition of viability of HL60 cells of tivantinib and its combination with either ABT-199 (left) or cytarabine (Ara-C; right). Synergy is assessed by the Bliss model of independence (histograms in insets). Displayed in the histograms are the experimentally determined differences for each drug combination from the calculated Bliss additivity on a scale of +20% to −20% cell viability in order of increasing tivantinib concentrations. Vertical lines indicate increments of 10% cell viability. Bars pointing up from the blue baseline (additivity) indicate synergy, bars pointing down indicate antagonism. ( c ) Effects of tivantinib and ABT-199 combination (in μM) on PARP-1 and caspase 3 cleavage as well as pSer10 histone H3 levels after 24 h treatment. ( d ) Effects of tivantinib and ABT-199 combination on β-catenin stabilization and pGSK3α/β Y279/216 levels. ( e ) Effects of tivantinib and ABT-199 combination on MCL-1, BCL-XL, and Bak. V = vehicle (DMSO). Tivantinib, ABT-199, and BIO concentrations are in μM. NaCl and LiCl concentrations are in mM.

    Article Snippet: Interestingly tivantinib single agent and, more pronouncedly, ABT-199 combination caused a loss of anti-apoptotic MCL-1 and BCL-XL protein levels while maintaining pro-apoptotic Bak levels (Fig. ).

    Techniques: Inhibition, Labeling, Acetylene Reduction Assay

    Proteomic analysis of tivantinib’s target profile. ( a ) Chemical structures of (−)-tivantinib and couplable c-(−)-tivantinib ( b ) Kinases enriched from drug affinity chromatography in HL60 cells passing SaintScore > 0.95, CRAPomePCT ≥ 95%, and -ln(NSAF) ≥ −7 cutoffs. Bubble size represents the sum of total unique spectra. Bubble color represents probability of a specific interaction based on the CRAPome ( c ) Total unique spectra of GSK3α and GSK3β for tivantinib and ampicillin control pulldowns. ( d ) Western blot of GSK3α and GKS3β following drug affinity chromatography experiments with c-(−)-tivantinib and c-(+)-tivantinib in HL60 and U937 cells. Competition experiments were performed with 20 μM BIO. TCL = total cell lysate, BB = blocked beads

    Journal: Scientific Reports

    Article Title: Off-target based drug repurposing opportunities for tivantinib in acute myeloid leukemia

    doi: 10.1038/s41598-018-37174-6

    Figure Lengend Snippet: Proteomic analysis of tivantinib’s target profile. ( a ) Chemical structures of (−)-tivantinib and couplable c-(−)-tivantinib ( b ) Kinases enriched from drug affinity chromatography in HL60 cells passing SaintScore > 0.95, CRAPomePCT ≥ 95%, and -ln(NSAF) ≥ −7 cutoffs. Bubble size represents the sum of total unique spectra. Bubble color represents probability of a specific interaction based on the CRAPome ( c ) Total unique spectra of GSK3α and GSK3β for tivantinib and ampicillin control pulldowns. ( d ) Western blot of GSK3α and GKS3β following drug affinity chromatography experiments with c-(−)-tivantinib and c-(+)-tivantinib in HL60 and U937 cells. Competition experiments were performed with 20 μM BIO. TCL = total cell lysate, BB = blocked beads

    Article Snippet: Interestingly tivantinib single agent and, more pronouncedly, ABT-199 combination caused a loss of anti-apoptotic MCL-1 and BCL-XL protein levels while maintaining pro-apoptotic Bak levels (Fig. ).

    Techniques: Affinity Chromatography, Western Blot

    Analysis of cellular response following tivantinib treatment. ( a ) Effects of (-)-tivantinib (in μM), NaCl (20 mM) and the pan-GSK3 inhibitor LiCl (20 mM) on β-catenin and pGSK3α/β Y279/216 levels in HL60 cells. ( b ) Effects of tivantinib (in μM), NaCl (20 mM), and LiCl (20 mM) on PARP-1 and caspase 3 cleavage as well as pSer10 histone H3 levels after 4 and 24 h. ( c ) Cell cycle analysis by DAPI DNA staining following treatment of HL60 cells with DMSO, NaCl (20 mM), LiCl (20 mM), or tivantinib (1 μM) for 24 h. ( d ) Analysis of early and late apoptotic populations by Annexin V staining following treatment of HL60 cells for 4, 12, 18, or 24 h with DMSO, tivantinib (in μM), NaCl (20 mM), or LiCl (20 mM). ( e ) Cellular differentiation of HL60 cells following treatment with DMSO, tivantinib, NaCl or LiCl for 72 and 96 h as assessed by CD11b staining. Asterisk denotes p

    Journal: Scientific Reports

    Article Title: Off-target based drug repurposing opportunities for tivantinib in acute myeloid leukemia

    doi: 10.1038/s41598-018-37174-6

    Figure Lengend Snippet: Analysis of cellular response following tivantinib treatment. ( a ) Effects of (-)-tivantinib (in μM), NaCl (20 mM) and the pan-GSK3 inhibitor LiCl (20 mM) on β-catenin and pGSK3α/β Y279/216 levels in HL60 cells. ( b ) Effects of tivantinib (in μM), NaCl (20 mM), and LiCl (20 mM) on PARP-1 and caspase 3 cleavage as well as pSer10 histone H3 levels after 4 and 24 h. ( c ) Cell cycle analysis by DAPI DNA staining following treatment of HL60 cells with DMSO, NaCl (20 mM), LiCl (20 mM), or tivantinib (1 μM) for 24 h. ( d ) Analysis of early and late apoptotic populations by Annexin V staining following treatment of HL60 cells for 4, 12, 18, or 24 h with DMSO, tivantinib (in μM), NaCl (20 mM), or LiCl (20 mM). ( e ) Cellular differentiation of HL60 cells following treatment with DMSO, tivantinib, NaCl or LiCl for 72 and 96 h as assessed by CD11b staining. Asterisk denotes p

    Article Snippet: Interestingly tivantinib single agent and, more pronouncedly, ABT-199 combination caused a loss of anti-apoptotic MCL-1 and BCL-XL protein levels while maintaining pro-apoptotic Bak levels (Fig. ).

    Techniques: Cell Cycle Assay, Staining, Cell Differentiation

    Effects of tivantinib on AML cell viability. ( a ) Correlation of predicted vs. actual area under the curve (AUC) values across all cell lines in the training and test sets. NRMSE = Normalized Root Mean Square Error. ( b ) Empirical cumulative distribution function (ECDF) comparing the predicted AML AUC values to all the non-AML AUC values in CTRPv2. Statistical significance was determined using a Kolmogorov-Smirnov test. ( c–d ) Dose response curves and IC 50 values for inhibition of viability by (−)-tivantinib, (+)-tivantinib, LiCl and PF-04217903 of ( c ) HL60 and ( d ) U937 cells following 72 h treatment. Displayed concentrations are in μM.

    Journal: Scientific Reports

    Article Title: Off-target based drug repurposing opportunities for tivantinib in acute myeloid leukemia

    doi: 10.1038/s41598-018-37174-6

    Figure Lengend Snippet: Effects of tivantinib on AML cell viability. ( a ) Correlation of predicted vs. actual area under the curve (AUC) values across all cell lines in the training and test sets. NRMSE = Normalized Root Mean Square Error. ( b ) Empirical cumulative distribution function (ECDF) comparing the predicted AML AUC values to all the non-AML AUC values in CTRPv2. Statistical significance was determined using a Kolmogorov-Smirnov test. ( c–d ) Dose response curves and IC 50 values for inhibition of viability by (−)-tivantinib, (+)-tivantinib, LiCl and PF-04217903 of ( c ) HL60 and ( d ) U937 cells following 72 h treatment. Displayed concentrations are in μM.

    Article Snippet: Interestingly tivantinib single agent and, more pronouncedly, ABT-199 combination caused a loss of anti-apoptotic MCL-1 and BCL-XL protein levels while maintaining pro-apoptotic Bak levels (Fig. ).

    Techniques: Inhibition

    Effect of tivantinib, ABT-199 and their combination on primary AML patient blasts. ( a ) Dotplot of relative primary AML BMNCs colony formation following treatment with tivantinib, ABT-199 or their combination for 14 or 19 days. Counts were averaged and normalized to DMSO. Patient mutational status for commonly altered genes is displayed. ( b ) Absolute primary AML blast colony count for patients 3 and 4 following treatment for 19 and 14 days, respectively. Synergy values (deviation from Bliss) are annotated. Combo = 1 μM tivantinib + 0.5 μM ABT-199.

    Journal: Scientific Reports

    Article Title: Off-target based drug repurposing opportunities for tivantinib in acute myeloid leukemia

    doi: 10.1038/s41598-018-37174-6

    Figure Lengend Snippet: Effect of tivantinib, ABT-199 and their combination on primary AML patient blasts. ( a ) Dotplot of relative primary AML BMNCs colony formation following treatment with tivantinib, ABT-199 or their combination for 14 or 19 days. Counts were averaged and normalized to DMSO. Patient mutational status for commonly altered genes is displayed. ( b ) Absolute primary AML blast colony count for patients 3 and 4 following treatment for 19 and 14 days, respectively. Synergy values (deviation from Bliss) are annotated. Combo = 1 μM tivantinib + 0.5 μM ABT-199.

    Article Snippet: Interestingly tivantinib single agent and, more pronouncedly, ABT-199 combination caused a loss of anti-apoptotic MCL-1 and BCL-XL protein levels while maintaining pro-apoptotic Bak levels (Fig. ).

    Techniques:

    The effect of treatment with SNDX-275 on Fas and FADD expression in LM7 lung metastases in vivo

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: The effect of treatment with SNDX-275 on Fas and FADD expression in LM7 lung metastases in vivo

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques: Expressing, In Vivo

    SNDX-275 increases expression of FADD protein in LM7 cells

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: SNDX-275 increases expression of FADD protein in LM7 cells

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques: Expressing

    Transfection of LM7 cells with the FDN plasmid induces expression of the inactive (truncated) form of FADD and reverses SNDX-275 induced sensitization of LM7 cells to FasL

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: Transfection of LM7 cells with the FDN plasmid induces expression of the inactive (truncated) form of FADD and reverses SNDX-275 induced sensitization of LM7 cells to FasL

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques: Transfection, Plasmid Preparation, Expressing

    Treatment with SNDX-275 increases the sensitivity of LM7 cells to FasL without increasing Fas receptor expression on the cell surface

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: Treatment with SNDX-275 increases the sensitivity of LM7 cells to FasL without increasing Fas receptor expression on the cell surface

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques: Expressing

    Treatment with SNDX-275 increases Fas expression on the surface of DLM8 cells and increases its sensitivity to FasL

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: Treatment with SNDX-275 increases Fas expression on the surface of DLM8 cells and increases its sensitivity to FasL

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques: Expressing

    Treatment with SNDX-275 increases acetylation of global H3 histones and on the fas gene promoter in DLM8 cells

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: Treatment with SNDX-275 increases acetylation of global H3 histones and on the fas gene promoter in DLM8 cells

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques:

    Treatment with SNDX-275 stimulate expression of Fas mRNA

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: Treatment with SNDX-275 stimulate expression of Fas mRNA

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques: Expressing

    Transfection of DLM8 cells with the FDN plasmid induces expression of the inactive (truncated) form of FADD and reverses SNDX-275 induced sensitization of DLM8 cells to FasL

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: Transfection of DLM8 cells with the FDN plasmid induces expression of the inactive (truncated) form of FADD and reverses SNDX-275 induced sensitization of DLM8 cells to FasL

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques: Transfection, Plasmid Preparation, Expressing

    Treatment with SNDX-275 changes the pattern of mRNA expression of the molecules downstream from Fas signaling in favor of apoptosis in LM7 cells

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: Treatment with SNDX-275 changes the pattern of mRNA expression of the molecules downstream from Fas signaling in favor of apoptosis in LM7 cells

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques: Expressing

    SNDX-275 increases expression of Fas mRNA in LM7 cells

    Journal:

    Article Title: Effect of the HDAC inhibitor SNDX-275 on Fas signaling in osteosarcoma cells and the feasibility of its topical application for the treatment of osteosarcoma lung metastases

    doi: 10.1002/cncr.25884

    Figure Lengend Snippet: SNDX-275 increases expression of Fas mRNA in LM7 cells

    Article Snippet: SNDX-275 agent was a kind gift from Syndax Pharmaceuticals Inc. (Waltham, MA).

    Techniques: Expressing

    Immunofluorescence histochemistry of the sections from the aortic arch apoE −/− mice on a high cholesterol diet. (a) IntegriSense, (b) ProSense, red: imaging agent, green: CD 68 antibody staining monocytes/macrophages, blue: DAPI staining of cell nucleus, yellow arrows: colocalization of agents and macrophages, red arrows: distribution of agents in the core of atherosclerotic lesion, Adv: adventitia.

    Journal: International Journal of Molecular Imaging

    Article Title: Quantitative Longitudinal Imaging of Vascular Inflammation and Treatment by Ezetimibe in apoE Mice by FMT Using New Optical Imaging Biomarkers of Cathepsin Activity and αvβ3 Integrin

    doi: 10.1155/2012/189254

    Figure Lengend Snippet: Immunofluorescence histochemistry of the sections from the aortic arch apoE −/− mice on a high cholesterol diet. (a) IntegriSense, (b) ProSense, red: imaging agent, green: CD 68 antibody staining monocytes/macrophages, blue: DAPI staining of cell nucleus, yellow arrows: colocalization of agents and macrophages, red arrows: distribution of agents in the core of atherosclerotic lesion, Adv: adventitia.

    Article Snippet: The cathepsin activated agent ProSense (Figures and ), used in the detection of atherosclerotic lesions before [ – ], is based on ~400 kDa polylysine macromolecules with covalently attached fluorophores [ ].

    Techniques: Immunofluorescence, Mouse Assay, Imaging, Staining

    Examples of in vivo FMT images of apoE −/− mice and ex vivo fluorescence images of the corresponding areas of dissected arteries and correlation between the in vivo and ex vivo measurements. (a), (b) ProSense ( r = 0.89, P

    Journal: International Journal of Molecular Imaging

    Article Title: Quantitative Longitudinal Imaging of Vascular Inflammation and Treatment by Ezetimibe in apoE Mice by FMT Using New Optical Imaging Biomarkers of Cathepsin Activity and αvβ3 Integrin

    doi: 10.1155/2012/189254

    Figure Lengend Snippet: Examples of in vivo FMT images of apoE −/− mice and ex vivo fluorescence images of the corresponding areas of dissected arteries and correlation between the in vivo and ex vivo measurements. (a), (b) ProSense ( r = 0.89, P

    Article Snippet: The cathepsin activated agent ProSense (Figures and ), used in the detection of atherosclerotic lesions before [ – ], is based on ~400 kDa polylysine macromolecules with covalently attached fluorophores [ ].

    Techniques: In Vivo, Mouse Assay, Ex Vivo, Fluorescence

    Quantitative in vivo FMT measurements of agents in the thorax area of apoE −/− mice ( n = 15 in each group, total 30 animals) on a high cholesterol and control C57BL/6 mice ( n = 9 for ProSense, n = 5 for IntegriSense study) on regular chow diet. (a) ProSense; (b) IntegriSense (mean ± SEM, * P

    Journal: International Journal of Molecular Imaging

    Article Title: Quantitative Longitudinal Imaging of Vascular Inflammation and Treatment by Ezetimibe in apoE Mice by FMT Using New Optical Imaging Biomarkers of Cathepsin Activity and αvβ3 Integrin

    doi: 10.1155/2012/189254

    Figure Lengend Snippet: Quantitative in vivo FMT measurements of agents in the thorax area of apoE −/− mice ( n = 15 in each group, total 30 animals) on a high cholesterol and control C57BL/6 mice ( n = 9 for ProSense, n = 5 for IntegriSense study) on regular chow diet. (a) ProSense; (b) IntegriSense (mean ± SEM, * P

    Article Snippet: The cathepsin activated agent ProSense (Figures and ), used in the detection of atherosclerotic lesions before [ – ], is based on ~400 kDa polylysine macromolecules with covalently attached fluorophores [ ].

    Techniques: In Vivo, Mouse Assay

    The effect of prophylactic treatment using ezetimibe on the FMT measured levels of agents in thorax areas of apoE −/− mice ( n = 15 in each group, total 120) on high cholesterol diet. (a) ProSense; (b) ProSense FAST; (c) Cat B FAST; (d) IntegriSense (mean ± SEM, * P

    Journal: International Journal of Molecular Imaging

    Article Title: Quantitative Longitudinal Imaging of Vascular Inflammation and Treatment by Ezetimibe in apoE Mice by FMT Using New Optical Imaging Biomarkers of Cathepsin Activity and αvβ3 Integrin

    doi: 10.1155/2012/189254

    Figure Lengend Snippet: The effect of prophylactic treatment using ezetimibe on the FMT measured levels of agents in thorax areas of apoE −/− mice ( n = 15 in each group, total 120) on high cholesterol diet. (a) ProSense; (b) ProSense FAST; (c) Cat B FAST; (d) IntegriSense (mean ± SEM, * P

    Article Snippet: The cathepsin activated agent ProSense (Figures and ), used in the detection of atherosclerotic lesions before [ – ], is based on ~400 kDa polylysine macromolecules with covalently attached fluorophores [ ].

    Techniques: Mouse Assay

    Basic characteristics of cathepsin and α v β 3 integrin agents. (a), (b) Cartoon and spectra of ProSense ~400 kDa polylysine-based [ 27 ]; (c), (d) Cartoon and spectra of FAST peptide-based agents ~23 kDa, PKM: pharmacokinetic modifier, absorption spectra (before activation—dash red and after activation: solid red), emission spectra: solid blue; (e), selectivity in cathepsin probe activation at 24 hours incubation; (f), Chemical structure of parent α v β 3 integrin ligand (top) and IntegriSense (bottom, ~1278 Da) NIRF: VivoTag fluorophore.

    Journal: International Journal of Molecular Imaging

    Article Title: Quantitative Longitudinal Imaging of Vascular Inflammation and Treatment by Ezetimibe in apoE Mice by FMT Using New Optical Imaging Biomarkers of Cathepsin Activity and αvβ3 Integrin

    doi: 10.1155/2012/189254

    Figure Lengend Snippet: Basic characteristics of cathepsin and α v β 3 integrin agents. (a), (b) Cartoon and spectra of ProSense ~400 kDa polylysine-based [ 27 ]; (c), (d) Cartoon and spectra of FAST peptide-based agents ~23 kDa, PKM: pharmacokinetic modifier, absorption spectra (before activation—dash red and after activation: solid red), emission spectra: solid blue; (e), selectivity in cathepsin probe activation at 24 hours incubation; (f), Chemical structure of parent α v β 3 integrin ligand (top) and IntegriSense (bottom, ~1278 Da) NIRF: VivoTag fluorophore.

    Article Snippet: The cathepsin activated agent ProSense (Figures and ), used in the detection of atherosclerotic lesions before [ – ], is based on ~400 kDa polylysine macromolecules with covalently attached fluorophores [ ].

    Techniques: Activation Assay, Incubation

    Correlation between quantitative immunohistochemical analysis of the macrophage content and the FMT measured agent amount in selected sections from apoE −/− mice on high cholesterol diet. (a) An example of dissected arteries from apoE −/− mice. (b) A mouse heart cartoon with six regions marked with red lines, where the arteries were sectioned for histology. (c) An example of stained section used in the macrophage quantification. (d), (e), (f) Plots of total agent amount measured in vivo by FMT as a function of macrophage count for ProSense ((d), r = 0.89, P

    Journal: International Journal of Molecular Imaging

    Article Title: Quantitative Longitudinal Imaging of Vascular Inflammation and Treatment by Ezetimibe in apoE Mice by FMT Using New Optical Imaging Biomarkers of Cathepsin Activity and αvβ3 Integrin

    doi: 10.1155/2012/189254

    Figure Lengend Snippet: Correlation between quantitative immunohistochemical analysis of the macrophage content and the FMT measured agent amount in selected sections from apoE −/− mice on high cholesterol diet. (a) An example of dissected arteries from apoE −/− mice. (b) A mouse heart cartoon with six regions marked with red lines, where the arteries were sectioned for histology. (c) An example of stained section used in the macrophage quantification. (d), (e), (f) Plots of total agent amount measured in vivo by FMT as a function of macrophage count for ProSense ((d), r = 0.89, P

    Article Snippet: The cathepsin activated agent ProSense (Figures and ), used in the detection of atherosclerotic lesions before [ – ], is based on ~400 kDa polylysine macromolecules with covalently attached fluorophores [ ].

    Techniques: Immunohistochemistry, Mouse Assay, Staining, In Vivo

    In vitro killing assay. Notes: Intracellular antifungal activity of the AmB–PGA formulation against Candida albicans inside RAW 264.7 cells. Macrophages were infected with C. albicans and treated with AmB–PGA nanoparticles. Percentage of infection was calculated by lysing macrophages (after 24 hours of treatment) with Triton X-100 (0.2%), subculturing in Sabouraud agar plates, and comparing with an untreated control. Experiments were carried out in triplicate. Each datum point represents the mean ± standard deviation. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; CFU, colony forming units; PGA, polyglutamic acid.

    Journal: International Journal of Nanomedicine

    Article Title: Self-assembled amphotericin B-loaded polyglutamic acid nanoparticles: preparation, characterization and in vitro potential against Candida albicans

    doi: 10.2147/IJN.S63155

    Figure Lengend Snippet: In vitro killing assay. Notes: Intracellular antifungal activity of the AmB–PGA formulation against Candida albicans inside RAW 264.7 cells. Macrophages were infected with C. albicans and treated with AmB–PGA nanoparticles. Percentage of infection was calculated by lysing macrophages (after 24 hours of treatment) with Triton X-100 (0.2%), subculturing in Sabouraud agar plates, and comparing with an untreated control. Experiments were carried out in triplicate. Each datum point represents the mean ± standard deviation. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; CFU, colony forming units; PGA, polyglutamic acid.

    Article Snippet: The antifungal agents AmBisome (Gilead Sciences Inc, San Dimas, CA, USA) and Fungizone (Apothecon, Princeton, NJ, USA) were reconstituted according to the manufacturers’ instructions.

    Techniques: In Vitro, Activity Assay, Infection, Subculturing Assay, Standard Deviation

    Absorption spectra of the AmB–PGA nanoparticle formulation and commercially available formulations of AmB (AmB-L, AmB-D) in ( A ) aqueous medium and ( B ) organic solvent. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; PGA, polyglutamic acid.

    Journal: International Journal of Nanomedicine

    Article Title: Self-assembled amphotericin B-loaded polyglutamic acid nanoparticles: preparation, characterization and in vitro potential against Candida albicans

    doi: 10.2147/IJN.S63155

    Figure Lengend Snippet: Absorption spectra of the AmB–PGA nanoparticle formulation and commercially available formulations of AmB (AmB-L, AmB-D) in ( A ) aqueous medium and ( B ) organic solvent. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; PGA, polyglutamic acid.

    Article Snippet: The antifungal agents AmBisome (Gilead Sciences Inc, San Dimas, CA, USA) and Fungizone (Apothecon, Princeton, NJ, USA) were reconstituted according to the manufacturers’ instructions.

    Techniques:

    Scanning electron micrographs of inhibition of Candida albicans biofilm mediated by the AmB–PGA formulation. Notes: ( A ) Untreated control image of biofilm, ( B ) AmB-L treated biofilm, ( C ) biofilm treated with AmB-D, and ( D ) biofilm treated with AmB–PGA. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; PGA, polyglutamic acid.

    Journal: International Journal of Nanomedicine

    Article Title: Self-assembled amphotericin B-loaded polyglutamic acid nanoparticles: preparation, characterization and in vitro potential against Candida albicans

    doi: 10.2147/IJN.S63155

    Figure Lengend Snippet: Scanning electron micrographs of inhibition of Candida albicans biofilm mediated by the AmB–PGA formulation. Notes: ( A ) Untreated control image of biofilm, ( B ) AmB-L treated biofilm, ( C ) biofilm treated with AmB-D, and ( D ) biofilm treated with AmB–PGA. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; PGA, polyglutamic acid.

    Article Snippet: The antifungal agents AmBisome (Gilead Sciences Inc, San Dimas, CA, USA) and Fungizone (Apothecon, Princeton, NJ, USA) were reconstituted according to the manufacturers’ instructions.

    Techniques: Inhibition

    In vitro cytotoxicity assay. Dose–response effects of AmB–PGA formulation on cytotoxicity against ( A ) KB cells and ( B ) RAW 264.7 cells. Notes: The cells were exposed for 24 hours to various AmB formulations at different drug concentrations. MTT values were normalized to the control cells. The data are reported as the mean ± standard deviation of four experiments. Free drug is pure AmB used in preparation of the complex. Abbreviations: AmB, amphotericin B; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PGA, polyglutamic acid; AmB-D, Fungizone ® ; AmB-L, Ambisome ® .

    Journal: International Journal of Nanomedicine

    Article Title: Self-assembled amphotericin B-loaded polyglutamic acid nanoparticles: preparation, characterization and in vitro potential against Candida albicans

    doi: 10.2147/IJN.S63155

    Figure Lengend Snippet: In vitro cytotoxicity assay. Dose–response effects of AmB–PGA formulation on cytotoxicity against ( A ) KB cells and ( B ) RAW 264.7 cells. Notes: The cells were exposed for 24 hours to various AmB formulations at different drug concentrations. MTT values were normalized to the control cells. The data are reported as the mean ± standard deviation of four experiments. Free drug is pure AmB used in preparation of the complex. Abbreviations: AmB, amphotericin B; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PGA, polyglutamic acid; AmB-D, Fungizone ® ; AmB-L, Ambisome ® .

    Article Snippet: The antifungal agents AmBisome (Gilead Sciences Inc, San Dimas, CA, USA) and Fungizone (Apothecon, Princeton, NJ, USA) were reconstituted according to the manufacturers’ instructions.

    Techniques: In Vitro, Cytotoxicity Assay, MTT Assay, Standard Deviation

    Semiquantitative measure of antibiofilm activity of PGA nanoparticles. Notes: Growth percentage was analyzed by comparing relative metabolic activity obtained by XTT metabolic assay taking the untreated control as 100%. The data represent the mean ± standard deviation of three determinants and are representative of three different experiments (ie, the experiment was done in triplicate) with similar observations. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; PGA, polyglutamic acid; XTT, 2, 3-bis(2-methoxy-4-nitro-5-sulfoph enyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide.

    Journal: International Journal of Nanomedicine

    Article Title: Self-assembled amphotericin B-loaded polyglutamic acid nanoparticles: preparation, characterization and in vitro potential against Candida albicans

    doi: 10.2147/IJN.S63155

    Figure Lengend Snippet: Semiquantitative measure of antibiofilm activity of PGA nanoparticles. Notes: Growth percentage was analyzed by comparing relative metabolic activity obtained by XTT metabolic assay taking the untreated control as 100%. The data represent the mean ± standard deviation of three determinants and are representative of three different experiments (ie, the experiment was done in triplicate) with similar observations. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; PGA, polyglutamic acid; XTT, 2, 3-bis(2-methoxy-4-nitro-5-sulfoph enyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide.

    Article Snippet: The antifungal agents AmBisome (Gilead Sciences Inc, San Dimas, CA, USA) and Fungizone (Apothecon, Princeton, NJ, USA) were reconstituted according to the manufacturers’ instructions.

    Techniques: Activity Assay, Metabolic Assay, Standard Deviation

    Hemolytic activity of AmB and AmB–PGA nanoparticles. The extent of damage caused to red blood cells by the AmB formulation was measured as percent lysis of total erythrocytes used in the individual sample. Notes: ( A ) Hemolysis caused by the AmB–PGA formulation after 1 hour of incubation with human red blood cells. ( B ) Hemolysis after a 24-hour incubation period. AmB-D, AmB-L and pure AmB used in preparation of the complex were used as controls. Data are pooled from three different experiments. Each datum point is a mean ± standard deviation. Abbreviations: AmB, amphotericin B; DMSO, dimethyl sulfoxide; PGA, polyglutamic acid; AmB-D, Fungizone ® ; AmB-L, Ambisome ® .

    Journal: International Journal of Nanomedicine

    Article Title: Self-assembled amphotericin B-loaded polyglutamic acid nanoparticles: preparation, characterization and in vitro potential against Candida albicans

    doi: 10.2147/IJN.S63155

    Figure Lengend Snippet: Hemolytic activity of AmB and AmB–PGA nanoparticles. The extent of damage caused to red blood cells by the AmB formulation was measured as percent lysis of total erythrocytes used in the individual sample. Notes: ( A ) Hemolysis caused by the AmB–PGA formulation after 1 hour of incubation with human red blood cells. ( B ) Hemolysis after a 24-hour incubation period. AmB-D, AmB-L and pure AmB used in preparation of the complex were used as controls. Data are pooled from three different experiments. Each datum point is a mean ± standard deviation. Abbreviations: AmB, amphotericin B; DMSO, dimethyl sulfoxide; PGA, polyglutamic acid; AmB-D, Fungizone ® ; AmB-L, Ambisome ® .

    Article Snippet: The antifungal agents AmBisome (Gilead Sciences Inc, San Dimas, CA, USA) and Fungizone (Apothecon, Princeton, NJ, USA) were reconstituted according to the manufacturers’ instructions.

    Techniques: Activity Assay, Lysis, Incubation, Standard Deviation

    Representative time-kill curve plot for Candida albicans in the presence of AmB-D, AmB-L, or AmB–PGA at 4× minimum inhibitory concentration. Notes: Wells containing no antibiotic were taken as the control. Assays were performed in quadruplicate. Each result is representative of at least three separate experiments. All values are shown as the mean ± standard deviation. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; CFU, colony forming units; PGA, polyglutamic acid.

    Journal: International Journal of Nanomedicine

    Article Title: Self-assembled amphotericin B-loaded polyglutamic acid nanoparticles: preparation, characterization and in vitro potential against Candida albicans

    doi: 10.2147/IJN.S63155

    Figure Lengend Snippet: Representative time-kill curve plot for Candida albicans in the presence of AmB-D, AmB-L, or AmB–PGA at 4× minimum inhibitory concentration. Notes: Wells containing no antibiotic were taken as the control. Assays were performed in quadruplicate. Each result is representative of at least three separate experiments. All values are shown as the mean ± standard deviation. Abbreviations: AmB, amphotericin B; AmB-D, Fungizone ® ; AmB-L, Ambisome ® ; CFU, colony forming units; PGA, polyglutamic acid.

    Article Snippet: The antifungal agents AmBisome (Gilead Sciences Inc, San Dimas, CA, USA) and Fungizone (Apothecon, Princeton, NJ, USA) were reconstituted according to the manufacturers’ instructions.

    Techniques: Concentration Assay, Standard Deviation

    Effect of MβCD on cell cycle and apoptosis of triple negative breast cancer cells. Cell cycle distribution of MDA-MB 231 (A) and MDA-MB 468 cells (B). Propidium iodide stained cells were analyzed for DNA content using flow cytometry. Histograms represent the percentage of MDA-MB 231 (C) and MDA-MB 468 (D) cells in G0/G1, S and G2/M phases. The data represent one of three independent experiments. Values are mean±SD of three different experiments. MDA-MB 231 (E) and MDA-MB 468 (F) cells were stained for apoptosis using TdT-mediated dUTP nick end-labeling (TUNEL) assay. Quantification of apoptotic cells expressed as a percent of 4′,6-diamidino-2-phenylindole (DAPI)-stained cells in MDA-MB231 (G) and MDA-MB 468 (H). Data shown from three independent experiments, bars represent the mean±SD of three experiments.

    Journal: Journal of Breast Cancer

    Article Title: Lipid Raft Integrity Is Required for Survival of Triple Negative Breast Cancer Cells

    doi: 10.4048/jbc.2016.19.4.372

    Figure Lengend Snippet: Effect of MβCD on cell cycle and apoptosis of triple negative breast cancer cells. Cell cycle distribution of MDA-MB 231 (A) and MDA-MB 468 cells (B). Propidium iodide stained cells were analyzed for DNA content using flow cytometry. Histograms represent the percentage of MDA-MB 231 (C) and MDA-MB 468 (D) cells in G0/G1, S and G2/M phases. The data represent one of three independent experiments. Values are mean±SD of three different experiments. MDA-MB 231 (E) and MDA-MB 468 (F) cells were stained for apoptosis using TdT-mediated dUTP nick end-labeling (TUNEL) assay. Quantification of apoptotic cells expressed as a percent of 4′,6-diamidino-2-phenylindole (DAPI)-stained cells in MDA-MB231 (G) and MDA-MB 468 (H). Data shown from three independent experiments, bars represent the mean±SD of three experiments.

    Article Snippet: TNBC cells were treated with the lipid raft disrupting agents MβCD, nystatin and filipin III (Sigma, St. Louis, USA) at concentrations of 0.1, 0.2, 0.3, 0.4, and 0.5 mM (0.1–0.5 mM) for 1, 24, and 48 hours.

    Techniques: Multiple Displacement Amplification, Staining, Flow Cytometry, Cytometry, End Labeling, TUNEL Assay

    Effect of cholesterol supplementation on survival and metastasis of methyl-β-cyclodextrin (MβCD) treated triple negative breast cancer cells. Cells were treated with MβCD for 48 hours and supplemented with cholesterol in the form of 1 mM MβCD-cholesterol complex for 24 hours. Proliferation of MDA-MB 231 cells (A) and MDA-MB 468 cells (B) in terms of cytotoxicity was measured using lactate dehydrogenase assay. Effect of cholesterol supplementation on adhesive potential of MDA-MB 231 and MDA-MB 468 (C) cells to fibronectin-coated and vitronectin-coated plates, respectively. Effect of cholesterol supplementation on Transwell invasion of MDA-MB 231 and 468 (D) cells through Matrigel and the percentage of MDA-MB 231 (E) and MDA-MB 468 (F) cells in G0/G1, S and G2/M phases. (G) Effect of cholesterol supplementation on TUNEL positive of MDA-MB231 and 468 cells. The values are expressed as means±SD of three independent experiments.

    Journal: Journal of Breast Cancer

    Article Title: Lipid Raft Integrity Is Required for Survival of Triple Negative Breast Cancer Cells

    doi: 10.4048/jbc.2016.19.4.372

    Figure Lengend Snippet: Effect of cholesterol supplementation on survival and metastasis of methyl-β-cyclodextrin (MβCD) treated triple negative breast cancer cells. Cells were treated with MβCD for 48 hours and supplemented with cholesterol in the form of 1 mM MβCD-cholesterol complex for 24 hours. Proliferation of MDA-MB 231 cells (A) and MDA-MB 468 cells (B) in terms of cytotoxicity was measured using lactate dehydrogenase assay. Effect of cholesterol supplementation on adhesive potential of MDA-MB 231 and MDA-MB 468 (C) cells to fibronectin-coated and vitronectin-coated plates, respectively. Effect of cholesterol supplementation on Transwell invasion of MDA-MB 231 and 468 (D) cells through Matrigel and the percentage of MDA-MB 231 (E) and MDA-MB 468 (F) cells in G0/G1, S and G2/M phases. (G) Effect of cholesterol supplementation on TUNEL positive of MDA-MB231 and 468 cells. The values are expressed as means±SD of three independent experiments.

    Article Snippet: TNBC cells were treated with the lipid raft disrupting agents MβCD, nystatin and filipin III (Sigma, St. Louis, USA) at concentrations of 0.1, 0.2, 0.3, 0.4, and 0.5 mM (0.1–0.5 mM) for 1, 24, and 48 hours.

    Techniques: Multiple Displacement Amplification, Lactate Dehydrogenase Assay, TUNEL Assay

    Effect of cholesterol depleting agents on cytotoxicity of triple negative breast cancer cells. MDA-MB 231 and 468 cells were treated with methyl-β-cyclodextrin (MβCD), nystatin and filipin III at different concentrations (0.1–0.5 mM) for 1, 24, and 48 hours and cell proliferation in terms of cytotoxicity was measured using lactate dehydrogenase assay. Cytotoxic effect of MβCD (A, B), nystatin (C, D) and filipin III (E, F) in MDA-MB 231 and MDA-MB 468 cells, respectively. The cytotoxicity was expressed as percent control. The results represent the mean±SD of three independent experiments.

    Journal: Journal of Breast Cancer

    Article Title: Lipid Raft Integrity Is Required for Survival of Triple Negative Breast Cancer Cells

    doi: 10.4048/jbc.2016.19.4.372

    Figure Lengend Snippet: Effect of cholesterol depleting agents on cytotoxicity of triple negative breast cancer cells. MDA-MB 231 and 468 cells were treated with methyl-β-cyclodextrin (MβCD), nystatin and filipin III at different concentrations (0.1–0.5 mM) for 1, 24, and 48 hours and cell proliferation in terms of cytotoxicity was measured using lactate dehydrogenase assay. Cytotoxic effect of MβCD (A, B), nystatin (C, D) and filipin III (E, F) in MDA-MB 231 and MDA-MB 468 cells, respectively. The cytotoxicity was expressed as percent control. The results represent the mean±SD of three independent experiments.

    Article Snippet: TNBC cells were treated with the lipid raft disrupting agents MβCD, nystatin and filipin III (Sigma, St. Louis, USA) at concentrations of 0.1, 0.2, 0.3, 0.4, and 0.5 mM (0.1–0.5 mM) for 1, 24, and 48 hours.

    Techniques: Multiple Displacement Amplification, Lactate Dehydrogenase Assay

    Effect of lipid rafts disruption on detergent resistant membrane (DRM) and non-DRM fractions. The isolated DRM and non-DRM fractions were subjected to marker specific enzyme-linked immunosorbent assay. Levels of caveolin-1 in DRM and non-DRM fractions of untreated and methyl-β-cyclodextrin (MβCD) treated cells of MDA-MB 231 (A) and MDA-MB 468 (B). The transferrin levels in DRM and non-DRM fractions of untreated and MβCD treated cells of MDA-MB 231 (C) and MDA-MB 468 (D).

    Journal: Journal of Breast Cancer

    Article Title: Lipid Raft Integrity Is Required for Survival of Triple Negative Breast Cancer Cells

    doi: 10.4048/jbc.2016.19.4.372

    Figure Lengend Snippet: Effect of lipid rafts disruption on detergent resistant membrane (DRM) and non-DRM fractions. The isolated DRM and non-DRM fractions were subjected to marker specific enzyme-linked immunosorbent assay. Levels of caveolin-1 in DRM and non-DRM fractions of untreated and methyl-β-cyclodextrin (MβCD) treated cells of MDA-MB 231 (A) and MDA-MB 468 (B). The transferrin levels in DRM and non-DRM fractions of untreated and MβCD treated cells of MDA-MB 231 (C) and MDA-MB 468 (D).

    Article Snippet: TNBC cells were treated with the lipid raft disrupting agents MβCD, nystatin and filipin III (Sigma, St. Louis, USA) at concentrations of 0.1, 0.2, 0.3, 0.4, and 0.5 mM (0.1–0.5 mM) for 1, 24, and 48 hours.

    Techniques: Isolation, Marker, Enzyme-linked Immunosorbent Assay, Multiple Displacement Amplification

    Effect of methyl-β-cyclodextrin (MβCD) on adhesion and invasion of triple negative breast cancer cells. (A) Adhesion assay was performed to evaluate the effects of MβCD on the adhesive potential of MDA-MB 231 and 468 cells to fibronectin and vitronectin-coated plates, respectively (stained with Hema 3, ×200). (B) Percent of adhesion was calculated from the mean obtained from three independent experiments and are represented (±SEM). (C) Transwell invasion assay was performed to evaluate the effects of MβCD on invasion of MDA-MB 231 and 468 cells through Matrigel (stained with Hema 3, ×200). (D) Percent of invasion was calculated from the mean obtained from three independent experiments and are represented (SEM).

    Journal: Journal of Breast Cancer

    Article Title: Lipid Raft Integrity Is Required for Survival of Triple Negative Breast Cancer Cells

    doi: 10.4048/jbc.2016.19.4.372

    Figure Lengend Snippet: Effect of methyl-β-cyclodextrin (MβCD) on adhesion and invasion of triple negative breast cancer cells. (A) Adhesion assay was performed to evaluate the effects of MβCD on the adhesive potential of MDA-MB 231 and 468 cells to fibronectin and vitronectin-coated plates, respectively (stained with Hema 3, ×200). (B) Percent of adhesion was calculated from the mean obtained from three independent experiments and are represented (±SEM). (C) Transwell invasion assay was performed to evaluate the effects of MβCD on invasion of MDA-MB 231 and 468 cells through Matrigel (stained with Hema 3, ×200). (D) Percent of invasion was calculated from the mean obtained from three independent experiments and are represented (SEM).

    Article Snippet: TNBC cells were treated with the lipid raft disrupting agents MβCD, nystatin and filipin III (Sigma, St. Louis, USA) at concentrations of 0.1, 0.2, 0.3, 0.4, and 0.5 mM (0.1–0.5 mM) for 1, 24, and 48 hours.

    Techniques: Cell Adhesion Assay, Multiple Displacement Amplification, Staining, Transwell Invasion Assay

    Effect of Lipid raft disruption on tumor induced angiogenesis and expression of angiogenic molecules. In vitro angiogenesis in MDA-MB 231 and MDA-MB 468 cells (A). Tumor-induced tube formation in human umbilical vascular endothelial cells (HUVEC) cells was carried out as described in METHODS. The tube formation was observed under the bright field microscope and number of branch points were calculated (stained with Hema 3, ×400). (B) Graphical representation of relative branch points in MDA-MB 231 and MDA-MB 468 cells treated with methyl-β-cyclodextrin (MβCD). Bars represents the mean±SE of three different experiments. Expression of pro and antiangiogenic molecules in HUVEC and MDA-MB 231 or MDA-MB 468 co-cultures. Conditioned media from HUVEC and MDA-MB 231 or MDA-MB 468 co-cultures, exposed to angiogenesis antibody arrays and processed as per manufacturer's instructions. Graphical representation of fold change of pro- and antiangiogenic molecules (C, D).

    Journal: Journal of Breast Cancer

    Article Title: Lipid Raft Integrity Is Required for Survival of Triple Negative Breast Cancer Cells

    doi: 10.4048/jbc.2016.19.4.372

    Figure Lengend Snippet: Effect of Lipid raft disruption on tumor induced angiogenesis and expression of angiogenic molecules. In vitro angiogenesis in MDA-MB 231 and MDA-MB 468 cells (A). Tumor-induced tube formation in human umbilical vascular endothelial cells (HUVEC) cells was carried out as described in METHODS. The tube formation was observed under the bright field microscope and number of branch points were calculated (stained with Hema 3, ×400). (B) Graphical representation of relative branch points in MDA-MB 231 and MDA-MB 468 cells treated with methyl-β-cyclodextrin (MβCD). Bars represents the mean±SE of three different experiments. Expression of pro and antiangiogenic molecules in HUVEC and MDA-MB 231 or MDA-MB 468 co-cultures. Conditioned media from HUVEC and MDA-MB 231 or MDA-MB 468 co-cultures, exposed to angiogenesis antibody arrays and processed as per manufacturer's instructions. Graphical representation of fold change of pro- and antiangiogenic molecules (C, D).

    Article Snippet: TNBC cells were treated with the lipid raft disrupting agents MβCD, nystatin and filipin III (Sigma, St. Louis, USA) at concentrations of 0.1, 0.2, 0.3, 0.4, and 0.5 mM (0.1–0.5 mM) for 1, 24, and 48 hours.

    Techniques: Expressing, In Vitro, Multiple Displacement Amplification, Microscopy, Staining

    Effect of cholesterol depleting agents on membrane cholesterol in triple negative breast cancer cells. MDA-MB 231 and 468 cells were treated with methyl-β-cyclodextrin (MβCD), nystatin and filipin III at different concentrations (0.1–0.5 mM) for 1, 24, and 48 hours and reduction in cellular cholesterol levels were measured. Reduction of cellular cholesterol with MβCD (A, B), nystatin (C, D) and filipin III (E, F) in MDA-MB 231 and MDA-MB 468 cells, respectively. The percent reduction in the cholesterol upon the treatments was calculated with respect to total cholesterol in the untreated cells, which was taken as 100%. The results represent the mean±SD of three independent experiments.

    Journal: Journal of Breast Cancer

    Article Title: Lipid Raft Integrity Is Required for Survival of Triple Negative Breast Cancer Cells

    doi: 10.4048/jbc.2016.19.4.372

    Figure Lengend Snippet: Effect of cholesterol depleting agents on membrane cholesterol in triple negative breast cancer cells. MDA-MB 231 and 468 cells were treated with methyl-β-cyclodextrin (MβCD), nystatin and filipin III at different concentrations (0.1–0.5 mM) for 1, 24, and 48 hours and reduction in cellular cholesterol levels were measured. Reduction of cellular cholesterol with MβCD (A, B), nystatin (C, D) and filipin III (E, F) in MDA-MB 231 and MDA-MB 468 cells, respectively. The percent reduction in the cholesterol upon the treatments was calculated with respect to total cholesterol in the untreated cells, which was taken as 100%. The results represent the mean±SD of three independent experiments.

    Article Snippet: TNBC cells were treated with the lipid raft disrupting agents MβCD, nystatin and filipin III (Sigma, St. Louis, USA) at concentrations of 0.1, 0.2, 0.3, 0.4, and 0.5 mM (0.1–0.5 mM) for 1, 24, and 48 hours.

    Techniques: Multiple Displacement Amplification

    Preclinical studies of motolimod (Moto) plus pegylated liposomal doxorubicin (PLD) in NSG-HIS mice. (A) NSG-HIS mice were given either PLD alone (intraperitoneally, 50 mg/m 2 ), motolimod alone (subcutaneously 2 days after PLD dosing, 1.5 mg/m 2 ), the combination,

    Journal:

    Article Title: Integrative Development of a TLR8 Agonist for Ovarian Cancer Chemo-immunotherapy

    doi: 10.1158/1078-0432.CCR-16-1453

    Figure Lengend Snippet: Preclinical studies of motolimod (Moto) plus pegylated liposomal doxorubicin (PLD) in NSG-HIS mice. (A) NSG-HIS mice were given either PLD alone (intraperitoneally, 50 mg/m 2 ), motolimod alone (subcutaneously 2 days after PLD dosing, 1.5 mg/m 2 ), the combination,

    Article Snippet: A standard dose-escalation design was used to evaluate the combination of 40 mg/m2 PLD with 3 dose levels of motolimod (2.5, 3.0, and 3.5 mg/m2 ), which were selected based on the results of a previous phase 1 dose-seeking study of single-agent motolimod (doses of 0.1 to 3.9 mg/m2 ) in adults with advanced cancer ( ).

    Techniques: Mouse Assay

    The NSG-HIS mouse model is suitable for the preclinical assessments of motolimod. (A) The in vivo plasma biomarker response to motolimod, administered at the respective doses shown, measured at 6 hours, is compared across NSG-HIS mice (n=3/group), non-human

    Journal:

    Article Title: Integrative Development of a TLR8 Agonist for Ovarian Cancer Chemo-immunotherapy

    doi: 10.1158/1078-0432.CCR-16-1453

    Figure Lengend Snippet: The NSG-HIS mouse model is suitable for the preclinical assessments of motolimod. (A) The in vivo plasma biomarker response to motolimod, administered at the respective doses shown, measured at 6 hours, is compared across NSG-HIS mice (n=3/group), non-human

    Article Snippet: A standard dose-escalation design was used to evaluate the combination of 40 mg/m2 PLD with 3 dose levels of motolimod (2.5, 3.0, and 3.5 mg/m2 ), which were selected based on the results of a previous phase 1 dose-seeking study of single-agent motolimod (doses of 0.1 to 3.9 mg/m2 ) in adults with advanced cancer ( ).

    Techniques: In Vivo, Biomarker Assay, Mouse Assay

    Biological effects of motolimod in humans. The in vivo cytokine response to motolimod was assessed in women with advanced ovarian cancer who were treated with 40 mg/m 2 of PLD in combination with motolimod at 2.5 (n=3), 3.0 (n=3), or 3.5 mg/m 2 (n=7). Plasma

    Journal:

    Article Title: Integrative Development of a TLR8 Agonist for Ovarian Cancer Chemo-immunotherapy

    doi: 10.1158/1078-0432.CCR-16-1453

    Figure Lengend Snippet: Biological effects of motolimod in humans. The in vivo cytokine response to motolimod was assessed in women with advanced ovarian cancer who were treated with 40 mg/m 2 of PLD in combination with motolimod at 2.5 (n=3), 3.0 (n=3), or 3.5 mg/m 2 (n=7). Plasma

    Article Snippet: A standard dose-escalation design was used to evaluate the combination of 40 mg/m2 PLD with 3 dose levels of motolimod (2.5, 3.0, and 3.5 mg/m2 ), which were selected based on the results of a previous phase 1 dose-seeking study of single-agent motolimod (doses of 0.1 to 3.9 mg/m2 ) in adults with advanced cancer ( ).

    Techniques: In Vivo

    Innate and adaptive immune-mediated interactions underlie the effect of the motolimod plus PLD combination. (A) OVCAR5 cells were evaluated in vitro for sensitivity to TNFα. Cells were left untreated (control) or were exposed to TNFα or

    Journal:

    Article Title: Integrative Development of a TLR8 Agonist for Ovarian Cancer Chemo-immunotherapy

    doi: 10.1158/1078-0432.CCR-16-1453

    Figure Lengend Snippet: Innate and adaptive immune-mediated interactions underlie the effect of the motolimod plus PLD combination. (A) OVCAR5 cells were evaluated in vitro for sensitivity to TNFα. Cells were left untreated (control) or were exposed to TNFα or

    Article Snippet: A standard dose-escalation design was used to evaluate the combination of 40 mg/m2 PLD with 3 dose levels of motolimod (2.5, 3.0, and 3.5 mg/m2 ), which were selected based on the results of a previous phase 1 dose-seeking study of single-agent motolimod (doses of 0.1 to 3.9 mg/m2 ) in adults with advanced cancer ( ).

    Techniques: In Vitro

    Translation of the PLD plus motolimod combination into the clinic

    Journal:

    Article Title: Integrative Development of a TLR8 Agonist for Ovarian Cancer Chemo-immunotherapy

    doi: 10.1158/1078-0432.CCR-16-1453

    Figure Lengend Snippet: Translation of the PLD plus motolimod combination into the clinic

    Article Snippet: A standard dose-escalation design was used to evaluate the combination of 40 mg/m2 PLD with 3 dose levels of motolimod (2.5, 3.0, and 3.5 mg/m2 ), which were selected based on the results of a previous phase 1 dose-seeking study of single-agent motolimod (doses of 0.1 to 3.9 mg/m2 ) in adults with advanced cancer ( ).

    Techniques:

    Motolimod induces human immune cell activation in the NSG-HIS mouse. (A) NSG-HIS mice (n=3 mice/group) were administered vehicle (control) or motolimod at 1.5 or 15 mg/m 2 . Plasma samples were obtained 6 hours after dosing and analyzed by HumanMAP ®

    Journal:

    Article Title: Integrative Development of a TLR8 Agonist for Ovarian Cancer Chemo-immunotherapy

    doi: 10.1158/1078-0432.CCR-16-1453

    Figure Lengend Snippet: Motolimod induces human immune cell activation in the NSG-HIS mouse. (A) NSG-HIS mice (n=3 mice/group) were administered vehicle (control) or motolimod at 1.5 or 15 mg/m 2 . Plasma samples were obtained 6 hours after dosing and analyzed by HumanMAP ®

    Article Snippet: A standard dose-escalation design was used to evaluate the combination of 40 mg/m2 PLD with 3 dose levels of motolimod (2.5, 3.0, and 3.5 mg/m2 ), which were selected based on the results of a previous phase 1 dose-seeking study of single-agent motolimod (doses of 0.1 to 3.9 mg/m2 ) in adults with advanced cancer ( ).

    Techniques: Activation Assay, Mouse Assay

    Phases and morphologies of the solid products from reaction of CuO NPs with soluble HS - . (A) XRD patterns of sulfidated CuO nanoparticles generated from initial Cu/S ratios that vary from 0.2 to 5. CuO (tenorite) and CuS (covellite) reference is presented

    Journal:

    Article Title: Biological and Environmental Transformations of Copper-Based Nanomaterials

    doi: 10.1021/nn403080y

    Figure Lengend Snippet: Phases and morphologies of the solid products from reaction of CuO NPs with soluble HS - . (A) XRD patterns of sulfidated CuO nanoparticles generated from initial Cu/S ratios that vary from 0.2 to 5. CuO (tenorite) and CuS (covellite) reference is presented

    Article Snippet: CuO NPs agent were purchased from Aldrich and used as received.

    Techniques: Generated

    Target cell uptake and toxicity of carbon black, CuS, and CuO NPs. (A) Confocal images of murine macrophages after exposure to 5 ppm of test particles for 3 hours; nuclei were visualized (blue fluorescence) using 4′6-diamidino-2-phenylindole (DAPI).

    Journal:

    Article Title: Biological and Environmental Transformations of Copper-Based Nanomaterials

    doi: 10.1021/nn403080y

    Figure Lengend Snippet: Target cell uptake and toxicity of carbon black, CuS, and CuO NPs. (A) Confocal images of murine macrophages after exposure to 5 ppm of test particles for 3 hours; nuclei were visualized (blue fluorescence) using 4′6-diamidino-2-phenylindole (DAPI).

    Article Snippet: CuO NPs agent were purchased from Aldrich and used as received.

    Techniques: Fluorescence

    Properties of sulfidated CuO NPs and implications for toxicity. (A,B) CuS clusters and nanoparticles serve as catalysts for bisulfide oxidation. Reaction tracked through depletion of dissolved oxygen upon addition of copper ions to excess Na 2 S solution.

    Journal:

    Article Title: Biological and Environmental Transformations of Copper-Based Nanomaterials

    doi: 10.1021/nn403080y

    Figure Lengend Snippet: Properties of sulfidated CuO NPs and implications for toxicity. (A,B) CuS clusters and nanoparticles serve as catalysts for bisulfide oxidation. Reaction tracked through depletion of dissolved oxygen upon addition of copper ions to excess Na 2 S solution.

    Article Snippet: CuO NPs agent were purchased from Aldrich and used as received.

    Techniques:

    (A) Generation of H 2 O 2 by murine macrophages exposed to carbon black, CuO and CuS NPs. Detection of H 2 O 2 production in macrophages exposed to 2.5 ppm, 5 ppm, or 10 ppm of CuO, CuS or M120 (carbon black) as determined using the Amplex Red assay 20 minutes

    Journal:

    Article Title: Biological and Environmental Transformations of Copper-Based Nanomaterials

    doi: 10.1021/nn403080y

    Figure Lengend Snippet: (A) Generation of H 2 O 2 by murine macrophages exposed to carbon black, CuO and CuS NPs. Detection of H 2 O 2 production in macrophages exposed to 2.5 ppm, 5 ppm, or 10 ppm of CuO, CuS or M120 (carbon black) as determined using the Amplex Red assay 20 minutes

    Article Snippet: CuO NPs agent were purchased from Aldrich and used as received.

    Techniques: Amplex Red Assay

    Dissolution behavior of CuO in various fluid media. (A) Soluble copper produced by 24 hr incubation of CuO NPs (initial concentration 200 ppm) as a function of pH (50 mM acetate buffers at pH 4, 5 and 6, PBS buffer at pH 7.4 and 50 mM borate buffers at

    Journal:

    Article Title: Biological and Environmental Transformations of Copper-Based Nanomaterials

    doi: 10.1021/nn403080y

    Figure Lengend Snippet: Dissolution behavior of CuO in various fluid media. (A) Soluble copper produced by 24 hr incubation of CuO NPs (initial concentration 200 ppm) as a function of pH (50 mM acetate buffers at pH 4, 5 and 6, PBS buffer at pH 7.4 and 50 mM borate buffers at

    Article Snippet: CuO NPs agent were purchased from Aldrich and used as received.

    Techniques: Produced, Incubation, Concentration Assay

    Hydroxyl radical EPR signal (DMPO spin trap) induced by (A) 2 or 20 ppm CuO NPs suspension or its filtrate containing only the soluble forms, (B) free copper ions (CuCl 2 ) at various concentrations (ppb ng-Cu/g-solution). These experiments used 1 mM hydrogen

    Journal:

    Article Title: Biological and Environmental Transformations of Copper-Based Nanomaterials

    doi: 10.1021/nn403080y

    Figure Lengend Snippet: Hydroxyl radical EPR signal (DMPO spin trap) induced by (A) 2 or 20 ppm CuO NPs suspension or its filtrate containing only the soluble forms, (B) free copper ions (CuCl 2 ) at various concentrations (ppb ng-Cu/g-solution). These experiments used 1 mM hydrogen

    Article Snippet: CuO NPs agent were purchased from Aldrich and used as received.

    Techniques: Electron Paramagnetic Resonance

    Ligand effects in the ROS activity of CuO-NPs and soluble salts. (A) Comparison of ROS activity of 20 ppm CuO NPs or its filtrate in cell culture media with 0.64 ppm copper ions in PBS buffer. Note that the ion released from 20 ppm CuO-NPs in medium is

    Journal:

    Article Title: Biological and Environmental Transformations of Copper-Based Nanomaterials

    doi: 10.1021/nn403080y

    Figure Lengend Snippet: Ligand effects in the ROS activity of CuO-NPs and soluble salts. (A) Comparison of ROS activity of 20 ppm CuO NPs or its filtrate in cell culture media with 0.64 ppm copper ions in PBS buffer. Note that the ion released from 20 ppm CuO-NPs in medium is

    Article Snippet: CuO NPs agent were purchased from Aldrich and used as received.

    Techniques: Activity Assay, Cell Culture

    (A) Optical images of concentrated secondary CuS particles through 200 nm filter (left) and the filtrate after CuO NPs suspension in NaOH solution through 200 nm filter (right). (B) UV-vis spectra shows formation of CuS nanoparticles. Curves 1,2,3 correspond

    Journal:

    Article Title: Biological and Environmental Transformations of Copper-Based Nanomaterials

    doi: 10.1021/nn403080y

    Figure Lengend Snippet: (A) Optical images of concentrated secondary CuS particles through 200 nm filter (left) and the filtrate after CuO NPs suspension in NaOH solution through 200 nm filter (right). (B) UV-vis spectra shows formation of CuS nanoparticles. Curves 1,2,3 correspond

    Article Snippet: CuO NPs agent were purchased from Aldrich and used as received.

    Techniques: