Journal: Scientific Reports

Article Title: Quinoline carboxylic acid derivatives as potent ectonucleotidase inhibitors

doi: 10.1038/s41598-026-36994-1

Figure Lengend Snippet: ( A ) Fluorescence spectroscopy and microscopy analysis of the antagonist drug molecule 4d. ( A ) Fluorescence spectroscopy profiles of 4d , excited between 410 and 500 nm, with corresponding emissions between 400 and 600 nm. ( B ) The bright-field image of A549 cells; Fluorescence microscopy image (red channel) of A549 cells treated with compound 4d; Merged image combining bright-field and fluorescence channels, highlighting the localization of the compound 4d on the cell membrane.

Article Snippet: Fluorescence imaging experiments were performed using A549 human lung carcinoma cells from American Type Cultures Collection (ATCC, Manassas, VA), while untreated A549 cells were used as controls in fluorescence imaging to account for background autofluorescence and confirm the specificity of compound 4d uptake and signal.

Techniques: Fluorescence, Spectroscopy, Microscopy, Membrane

Journal:

Article Title: UXT Is a Novel Centrosomal Protein Essential for Cell Viability D⃞

doi: 10.1091/mbc.E05-08-0705

Figure Lengend Snippet: UXT is a novel centrosomal component. (A) Colocalization of UXT with the centrosome marker protein γ-tubulin. U2OS cells stably expressing EGFP:UXT (green) were labeled with anti-γ-tubulin (red). Images of cells in the G1, S/G2, prometaphase, metaphase, and anaphase are shown. (B) GFP-UXT is localized to the spindle poles during mitosis. U2OS cells stably expressing EGFP:UXT (green) were stained with an anti-α-tubulin (red) antibody. (C) The FLAG:UXT protein is localized to the centrosome. U2OS cells transfected with FLAG:UXT were immunostained with the mouse anti-FLAG antibody (red) and the rabbit anti-γ-tubulin antibody (green).

Article Snippet: Cell Culture and Treatments The human osteoblast sarcoma U2 (U2OS) cell line and the human embryonic kidney 293T (HEK293T) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Marker, Stable Transfection, Expressing, Labeling, Staining, Transfection

Journal:

Article Title: UXT Is a Novel Centrosomal Protein Essential for Cell Viability D⃞

doi: 10.1091/mbc.E05-08-0705

Figure Lengend Snippet: Characterization of the anti-UXT antibodies. (A) The anti-UXT antibody 1B2 can detect both the endogenous UXT and the UXT fusion proteins. Cell lysates from HEK293T cell (lane 1), HEK293T cells transfected with EGFP:UXT (lane 2), and HEK293T cells transfected with FLAG-tagged UXT (lane 3) were separated by 15% SDS-PAGE. The expression of UXT was examined with the anti-UXT antibody 1B2 by Western blot analysis. Note that the anti-UXT antibody 1B2 can detect the endogenous UXT protein (18KD), EGFP:UXT (50KD), and FLAG:UXT (25KD). (B and C) Immunofluorescence detection of the endogenous UXT proteins in U2OS cells using the anti-UXT antibodies. U2OS cells were fixed with methanol and stained with anti-γ-tubulin antibody (green) in combination with either the anti-UXT antibody 15A6 (B, in red) or the anti-UXT antibody 6D3 (C, in red). DNA was stained with DAPI (blue). The arrows indicate the centrosomes.

Article Snippet: Cell Culture and Treatments The human osteoblast sarcoma U2 (U2OS) cell line and the human embryonic kidney 293T (HEK293T) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Transfection, SDS Page, Expressing, Western Blot, Immunofluorescence, Staining

Journal:

Article Title: UXT Is a Novel Centrosomal Protein Essential for Cell Viability D⃞

doi: 10.1091/mbc.E05-08-0705

Figure Lengend Snippet: Overexpression of UXT disrupts the centrosome structure. (A) Loss of centrosomal γ-tubulin staining in U2OS cells after overexpression of EGFP:UXT. U2OS cells with transient expression of EGFP:UXT (green) were immunostained with the anti-γ-tubulin antibody (red). DNA was stained with DAPI. Note that the γ-tubulin staining on the centrosome is diminished in cells with over expression of GFP-UXT. (B) Electron microscopic image of disorganized centrosome in U2OS cells with overexpression of EGFP: UXT. U2OS cells, transfected with either EGFP or GFP-UXT, were processed for electron microscope imaging. Top, image of the centrosome in U2OS cells transfected with EGFP (control). Bottom, image of the abnormal centrosome in U2OS cells transfected with EGFP:UXT. The arrow indicates a normal centriole.

Article Snippet: Cell Culture and Treatments The human osteoblast sarcoma U2 (U2OS) cell line and the human embryonic kidney 293T (HEK293T) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Over Expression, Staining, Expressing, Transfection, Microscopy, Imaging, Control

Journal:

Article Title: UXT Is a Novel Centrosomal Protein Essential for Cell Viability D⃞

doi: 10.1091/mbc.E05-08-0705

Figure Lengend Snippet: UXT siRNA knockdown causes cell death. Two nonoverlapping UXT siRNAs were tested and produced similar results. The data obtained with one UXT siRNA were shown. The data obtained with another UXT siRNA were shown as supplemental data (Figure S2). (A) Efficacy of siRNA knockdown of the endogenous UXT protein. U2OS cells were treated with transfection reagent (NT), nonspecific siRNA (NS siRNA), or siRNA for UXT (UXT siRNA), respectively. The cell lysates were subject to Western blot using either the anti-UXT antibody 1B2 (top) or the anti-β-tubulin antibody (bottom). The protein levels of UXT were specifically reduced after 72-h treatment with siRNA for UXT. (B) UXT knockdown inhibits cell proliferation. U2OS cells were treated with nonspecific siRNA (left) or siRNA for UXT (right). Pictures were taken 72 h later. (C) UXT knockdown leads to cell death. U2OS cells were treated with nonspecific RNA or the UXT siRNA oligos. Seventy-two hours later, all the cells were collected and used for propidium iodide staining of DNA content by fluorescence-activated cell sorting. (D) p53 is not required for cell death caused by UXT knockdown. The HCT116 (p53+/+) or p53-negative HCT116 (p53-/-) cells were treated with either the control siRNA or the UXT siRNA. Seventy-two hours after transfection, the percentage of cell death was assessed by the trypan blue assay. The diagram shows the representative results of three experiments.

Article Snippet: Cell Culture and Treatments The human osteoblast sarcoma U2 (U2OS) cell line and the human embryonic kidney 293T (HEK293T) cell line were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Knockdown, Produced, Transfection, Western Blot, Staining, Fluorescence, FACS, Control

Journal:

Article Title: The Long Terminal Repeat of Jaagsiekte Sheep Retrovirus Is Preferentially Active in Differentiated Epithelial Cells of the Lungs

doi:

Figure Lengend Snippet: Cell lines used in this study

Article Snippet: H441 and H358 cells were grown in RPMI 1640 medium (Gibco BRL) adjusted to contain 1.5 g of sodium bicarbonate per liter, 4.5 g of glucose per liter, and 10 mM HEPES with 10% FBS. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Cell line Origin Tissue or cell type ATCC no. or reference A549 Human Lung carcinoma CCL-185 H441 Human BAC HTB-174 H358 Human BAC CRL-5807 293T Human Embryonic kidney Lebkowsky et al. ( 33 ) OA1 Sheep Brain fibroblast CRL-6538 OAT Sheep Sheep testis CRL-6546 FLL Sheep Primary fetal lamb MRI a JS7 Sheep BAC Jassim ( 30 ) CP-MRI Sheep Choroid plexus MRI a CP-ATCC Sheep Choroid plexus CRL-1700 mtCC1-2 Mouse Clara cell Magdaleno et al. ( 34 ) BV2 Mouse Microglia A. Tenner a F9 Mouse Testicular carcinoma CRL-1720 FOP Mouse Mammary carcinoma J. Hassel a IC-21 Mouse Peritoneal macrophage TIB-186 MHS Mouse Alveolar macrophage CRL-2019 C2C12 Mouse Myoblast CRL-1772 ABI-2 Mouse Hybridoma HB-33 MLE-12 Mouse Lung epithelium CRL-2110 TCMK Mouse Mouse kidney CCL-139 NIH 3T3 Mouse Mouse embryo CCL-92 MLE-15 Mouse Type II pneumocyte Wikenheiser et al. ( 59 ) ST3 Mouse Thymus stroma Brightman et al. ( 10 ) Open in a separate window a Cells were provided directly by an investigator or institution with no reference available.

Techniques:

Journal: European Journal of Nuclear Medicine and Molecular Imaging

Article Title: ApoSense : a novel technology for functional molecular imaging of cell death in models of acute renal tubular necrosis

doi: 10.1007/s00259-005-1905-x

Figure Lengend Snippet: Concomitant confocal imaging of HeLa cell undergoing apoptosis by DDC (green fluorescence) and annexin V (red fluorescence). DDC accumulates within the cytoplasm of the apoptotic cell while annexin V is attached to the external membrane

Article Snippet: Human adult T-cell leukemia Jurkat cells (clone E6-1) and cervical carcinoma HeLa cells (CCL-2.2), were purchased from ATCC (Rockville, MD, USA) and grown in Dulbecco’s modified Eagle’s medium with 10% fetal calf serum at 37°C in 5% CO 2 .

Techniques: Imaging, Fluorescence, Membrane

Journal: Oncotarget

Article Title: Diphenyl ditelluride anticancer activity and DNA topoisomerase I poisoning in human colon cancer HCT116 cells

doi: 10.18632/oncotarget.28465

Figure Lengend Snippet: ( A ) HCT116 and MRC5 cells were treated for 72 hours with increasing concentrations of DPDT (range: 0.1-20 μM) and cell viability assessed by the MTT assay. ( B ) HCT116 and MRC5 cells were treated for 72 hours with increasing concentrations of DPDT (range: 0.1–3 μM) and clone survival assessed by the colony-formation assay. All values are averages of at least 3 independent experiments done in duplicate. P values relative to the untreated control cells were calculated using one-way ANOVA Dunnett’s multiple comparison test: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human fetal lung fibroblast (MRC5) and colon carcinoma (HCT116) cells were purchased from American Type Culture Collection (ATCC).

Techniques: MTT Assay, Colony Assay, Control, Comparison

Journal: Oncotarget

Article Title: Diphenyl ditelluride anticancer activity and DNA topoisomerase I poisoning in human colon cancer HCT116 cells

doi: 10.18632/oncotarget.28465

Figure Lengend Snippet: ( A ) Representative flow cytometry diagrams obtained for HCT116 cells treated or not with DPDT. HCT116 cells were left untreated or incubated with 5 or 10 μM DPDT for 48 hours. The percentage of cells in Q4 (living cells, annexin V and PI negative cells), Q3 (early apoptotic cells, annexin V positive and PI negative cells) and Q2 (late apoptotic cells, annexin V and PI positive cells) was determined by flow cytometry. ( B ) Representative flow cytometry diagrams obtained for HCT116 cells treated or not with DPDT for 48 hours in presence of the pan-caspase inhibitor QVD-Oph (10 μM). ( C ) Data are expressed as the mean +/− SD ( n ≥ 3). Abbreviations: LC: Living cells; EA: Early apoptotic cells; LA: Late apoptotic cells. SDs are indicated by error bars when they exceed symbol size. P values relative to the QVD-Oph untreated cells were calculated using one-way ANOVA Dunnett's multiple comparison test: * p < 0.05.

Article Snippet: Human fetal lung fibroblast (MRC5) and colon carcinoma (HCT116) cells were purchased from American Type Culture Collection (ATCC).

Techniques: Flow Cytometry, Incubation, Comparison

Journal: Oncotarget

Article Title: Diphenyl ditelluride anticancer activity and DNA topoisomerase I poisoning in human colon cancer HCT116 cells

doi: 10.18632/oncotarget.28465

Figure Lengend Snippet: ( A and B ) show imaging of either MRC5 ( A ) or HCT116 ( B ) cells mounted on low melting agarose pre-coated slides after treatment with 0, 5 or 10 μM DPDT for 3 hours. Treatment with 1 μM CPT served as positive control. After drug exposure, cells were labeled with an anti-topoisomerase I antibody and nuclei were counterstained with DAPI. ( C ) Fluorescence intensities were quantified and data are expressed as the mean +/− SD ( n ≥ 3). P values were calculated using One Way ANOVA Dunnett's multiple comparison test ( ** p < 0.01; **** p < 0.0001).

Article Snippet: Human fetal lung fibroblast (MRC5) and colon carcinoma (HCT116) cells were purchased from American Type Culture Collection (ATCC).

Techniques: Imaging, Positive Control, Labeling, Fluorescence, Comparison

Journal: Oncotarget

Article Title: Diphenyl ditelluride anticancer activity and DNA topoisomerase I poisoning in human colon cancer HCT116 cells

doi: 10.18632/oncotarget.28465

Figure Lengend Snippet: Proliferating MRC5 and HCT116 cells were treated for 3 hours ( A ) or 24 hours ( B ) with 0, 1, 5, and 10 μM DPDT or 40 μM MMS (3 h) and subjected to alkaline comet assay. ( C ) Proliferating HCT116 cells were incubated with EdU for 30 min followed by 1 hour exposure to 0, 10 or 100 μM DPDT or 100 nM SN-38. Cells were fixed and processed for EdU and γ-H2AX staining, and the DNA was counterstained with Hoechst. ( D ) Increased magnification of the 10 μM DPDT treated cells shown in (C). The merged image illustrates the co-localization of EdU and γ-H2AX (indicated with arrows). ( E ) HCT116 cells were mock-treated or exposed for 1 hour to DPDT (10 or 100 μM) or SN-38 (100 nM). Cells were then fixed and processed for immunolabeling with an antibody directed against γ-H2AX. The fluorescence intensities were quantified and are indicated in arbitrary units (a.u.). At least 100 cells were analyzed for each condition. P values relative to the untreated cells were calculated using one-way ANOVA with Dunnett’s multiple comparison test: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human fetal lung fibroblast (MRC5) and colon carcinoma (HCT116) cells were purchased from American Type Culture Collection (ATCC).

Techniques: Alkaline Single Cell Gel Electrophoresis, Incubation, Staining, Immunolabeling, Fluorescence, Comparison

Journal: Oncotarget

Article Title: Diphenyl ditelluride anticancer activity and DNA topoisomerase I poisoning in human colon cancer HCT116 cells

doi: 10.18632/oncotarget.28465

Figure Lengend Snippet: Proliferating MRC5 and HCT116 cells were treated for 3 hours ( A ) or 24 hours ( B ) with 0, 1, 5, and 10 μM DPDT or 100 μM t-BOOH and subjected to DCFH-DA fluorescent labeling. The fluorescence intensities were quantified by flow cytometry and are expressed as the percentage of fluorescence increase in relation to the mock-treated cells (mean ± SD), n = 3. P values relative to the untreated cells were calculated using one-way ANOVA with Dunnett’s multiple comparison test: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human fetal lung fibroblast (MRC5) and colon carcinoma (HCT116) cells were purchased from American Type Culture Collection (ATCC).

Techniques: Labeling, Fluorescence, Flow Cytometry, Comparison

Journal: Pharmaceutics

Article Title: NIR Stimulus-Responsive PdPt Bimetallic Nanoparticles for Drug Delivery and Chemo-Photothermal Therapy

doi: 10.3390/pharmaceutics12070675

Figure Lengend Snippet: DOX release behaviors and in vitro cytotoxicity of DOX@PdPt@HA NPs. ( A ) NIR laser irradiation triggers release of DOX from DOX@PdPt@HA NPs. ( B ) Cell viability of 4T1 cells treated with different concentration of PdPt@HA NPs, DOX@PdPt@HA NPs or free DOX with or without NIR laser irradiation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. ( C , D ) Representative flow cytometry scatter plots of Annexin V/PI and quantification of apoptotic ratio of 4T1 cells with different treatments. ( E ) Fluorescence images of 4T1 cells exposed to the NPs with and without pre-incubation of hyaluronic acid (HA) for 1 h. Value represent mean ± SE of 3 independent experiments. P values: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Mouse mammary carcinoma 4T1 cells (Cell Bank of the Chinese Academy of Sciences, Shanghai, China) and human foreskin fibroblast (HFF; ATCC, Manassas, VA) cells were cultured in DMEM complete medium containing 100 U mL −1 penicillin, 100 U mL −1 streptomycin and 10% FBS.

Techniques: In Vitro, Irradiation, Concentration Assay, MTT Assay, Flow Cytometry, Fluorescence, Incubation

Journal: Pharmaceutics

Article Title: NIR Stimulus-Responsive PdPt Bimetallic Nanoparticles for Drug Delivery and Chemo-Photothermal Therapy

doi: 10.3390/pharmaceutics12070675

Figure Lengend Snippet: DOX@PdPt@HA NPs for in vivo combination therapy. ( A ) Thermal images of 4T1 tumor-bearing mice recorded by infrared thermal imaging camera. ( B ) Temperature changes of 4T1 tumors in different groups during laser irradiation. ( C , D ) Body weight of mice and growth of 4T1 tumors were monitored during the in vivo study. ( E ) Representative photographs of mice and excised tumors at the end of study. ( F ) Weight of tumor collected from animals in different groups. ns, no significant difference. Values represent mean ± SE ( n = 6 in each group). P values: ** P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Mouse mammary carcinoma 4T1 cells (Cell Bank of the Chinese Academy of Sciences, Shanghai, China) and human foreskin fibroblast (HFF; ATCC, Manassas, VA) cells were cultured in DMEM complete medium containing 100 U mL −1 penicillin, 100 U mL −1 streptomycin and 10% FBS.

Techniques: In Vivo, Imaging, Irradiation