Journal: Journal of Hepatocellular Carcinoma

Article Title: Coilin Affects the Prognosis of Hepatocellular Carcinoma Through Cell Cycle and Apoptosis

doi: 10.2147/JHC.S500119

Figure Lengend Snippet: Expression of COIL in HCC cell lines and HCC tissue from hospital. The expression of COIL in liver cancer cell lines huh7, hepG2 and hep3B was higher than that in L02 by immunohistochemistry ( A ) and Western blotting ( B ). ( C ) The relative expression of COIL in HCC was significantly higher than that in normal liver tissue by qRT-PCR (***p value < 0.001). ( D ) The expression of COIL in HCC tissue was higher than that in normal liver tissue. ( E, F ) Representative immunohistochemistry results of low or high COIL expression. ( G ) High COIL expression was associated with poor overall survival.

Article Snippet: The human normal liver cell line L02 was obtained from Beijing Baiao Bowei Biotechnology Co., Ltd.

Techniques: Expressing, Immunohistochemistry, Western Blot, Quantitative RT-PCR

Journal: Developmental cell

Article Title: Single-Cell and Population-Level Analyses Using Real-Time Kinetic Labeling Couples Proliferation and Cell Death Mechanisms

doi: 10.1016/j.devcel.2019.08.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, Peptides, and Recombinant Proteins ABT-737 Selleck Chemicals Cat#: S1002 mTNFα Peprotech Cat#: 315-01A mTRAIL Peprotech Cat#: 315-19 STYO21 Thermo Fisher Scientific Cat#: S7556 YOYO-3 Iodide Thermo Fisher Scientific Cat#: Y3606 zVAD-fmk APExBIO Cat#: A1902 Experimental Models: Cell Lines Bax +/+ Bak +/+ SV40 transformed MEF ATCC Cat#: CRL-2907; RRID: CVCL_U630 Bax −/− Bak −/− SV40 transformed MEF ATCC Cat#: CRL-2913; RRID: CVCL_U626 Cyld −/− SV40 transformed MEF Adrian Ting (Icahn School of Medicine at Mount Sinai, NY, USA) O’Donnell et al., 2011 Mlkl −/− adult ear fibroblasts Warren Alexander (University of Melbourne, Melbourne, Australia) Murphy et al., 2013 Recombinant DNA pProEx.Htb.annexinV Seamus Martin (Trinity College, Dublin, Ireland) Accession#: {"type":"entrez-nucleotide","attrs":{"text":"NM_001154","term_id":"1519243621","term_text":"NM_001154"}} NM_001154 ; Logue et al., 2009 Software and Algorithms Excel v16.16.9 Microsoft N/A ImageJ/Fiji v2.0.0-rc-69/1.52n National Institutes of Health N/A IncuCyte ZOOM v2018A Essen Biosciences N/A Prism v8.1.1 Graphpad Software N/A Sequential labeling pipeline This paper https://doi.org/10.5281/zenodo.3458574 Open in a separate window KEY RESOURCES TABLE SPARKL workflows use live-cell imagers to capture the kinetics of cell death in real time Multi-parametric analyses of cell death kinetics reveal mechanisms for comparative study Multiplex workflows accounts for cell-inherent or drug-induced proliferation changes Single-cell analyses quantify differential response to drugs in isogenic cell populations

Techniques: Recombinant, Transformation Assay, Software, Labeling

Journal: International Journal of Nanomedicine

Article Title: Differences in the Cell Type-Specific Toxicity of Diamond Nanoparticles to Endothelial Cells Depending on the Exposure of the Cells to Nanoparticles

doi: 10.2147/IJN.S411424

Figure Lengend Snippet: Diamond nanoparticles are cytotoxic to HUVEC but not to HS-5 and HMEC. HUVEC, HS-5, and HMEC 48-h proliferation ( A ) and 24-h membrane perforation ( B ) after treatment with NDs at concentrations of 5, 10, 20, 50, and 100 μg/mL. Statistical significance is indicated with asterisks: *P < 0.033, **P < 0.002, ***P < 0.001 (multifactor ANOVA; P < 0.05; n = 3 with 4 individual replicates for proliferation and 6 individual replicates for membrane perforation). All values are expressed as mean ± standard deviation. ( C ) HUVEC metabolic activity after treatment with NDs at concentrations of 5, 10, 20, 50, and 100 mg/l for 24 h. Statistical significance is indicated with different superscripts: P<0.001 (a, b, c, d, e) (ANOVA; P < 0.05; n = 3 with 6 individual replicates). ( D ) Live/Dead assay confocal microscopy images of nontreated HUVEC cells (“C”) and treated cells with 50 mg/l for 24 h (“ND”). The nuclei of dead cells are labelled with a green stain (NucGreen Dead 488), whereas all nuclei are stained with a red stain (NucRed Live 647). Images taken using Nomarski interference contrast (“NOM”) showed light-reflecting nanoparticle agglomerates, suggesting strong endocytosis of NDs.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) and human mammary epithelial cells (HMEC) were obtained from Thermo Fisher Scientific, whereas HS-5 cells were obtained from ATCC (LGC Standards, Łomianki, Poland).

Techniques: Membrane, Standard Deviation, Activity Assay, Live Dead Assay, Confocal Microscopy, Staining

Journal: International Journal of Nanomedicine

Article Title: Differences in the Cell Type-Specific Toxicity of Diamond Nanoparticles to Endothelial Cells Depending on the Exposure of the Cells to Nanoparticles

doi: 10.2147/IJN.S411424

Figure Lengend Snippet: Toxicity of diamond nanoparticles to HUVEC depends on cell density and surface conjugation of nanoparticles. ( A ) HUVEC 48-h proliferation was analysed for untreated cells (“C”) or cells treated with NDs (“ND”, “-RGD -FBS”) and NDs conjugated with a peptide containing an RGD motive (Gly-Arg-Gly-Asp-Ser; “ND”, “+RGD”) and at concentrations of 5 and 50 mg/l in standard HUVEC culture medium and in medium with the addition of 10% FBS (“+FBS”). Statistical significance is indicated with different superscripts: P<0.001 (a, b, c, d, e) (ANOVA; P < 0.05; n = 3 with 4 individual replicates). ( B ) Dependence of ND toxicity on HUVEC cell density was assessed in a 96-well plate at densities of 5 x 10 3 , 1 x 10 4 , 1.5 x 10 4 , and 2×10 4 . After 24-h incubation, cells were treated with NDs at a concentration of 0.4 ng/cell (the final ND concentration was 20, 40, 60, and 80 mg/L in 100 µL medium, respectively), and the metabolic activity was assessed. The relative toxicity of HUVEC was expressed as the ratio of the metabolic activity value of the ND-treated cells to the metabolic activity of the untreated cells. Statistical significance is indicated with different superscripts: P<0.001 (a, b, c, d) (ANOVA; P < 0.05; n = 3 with 6 individual replicates). All values are expressed as mean ± standard deviation. ( C ) HUVEC time-lapse images of a 12-h cell culture with NDs at a final concentration of 20 mg/l. The full video of time-lapse images is available as Supplementary Video . Abbreviations: RV, relative value; FUs, fluorescent units; C, control; NDs, diamond nanoparticles; RGD, Gly-Arg-Gly-Asp-Ser peptide; FBS, fetal bovine serum. ( D ) HUVEC spheroid morphology after 24-h treatment with NDs at a concentration of 10 and 100 mg/l. HUVEC spheroid membrane perforation ( E ) and spheroid size ( F ) for a after 24-h incubation with NDs at a concentration of 5, 10, 20, 50, and 100 mg/l. Statistical significance is indicated with different superscripts:: P<0.001 (a, b, c) (ANOVA; P < 0.05; n = 3 with 6 individual replicates).

Article Snippet: Human umbilical vein endothelial cells (HUVEC) and human mammary epithelial cells (HMEC) were obtained from Thermo Fisher Scientific, whereas HS-5 cells were obtained from ATCC (LGC Standards, Łomianki, Poland).

Techniques: Conjugation Assay, Incubation, Concentration Assay, Activity Assay, Standard Deviation, Cell Culture, Control, Membrane

Journal: International Journal of Nanomedicine

Article Title: Differences in the Cell Type-Specific Toxicity of Diamond Nanoparticles to Endothelial Cells Depending on the Exposure of the Cells to Nanoparticles

doi: 10.2147/IJN.S411424

Figure Lengend Snippet: Diamond nanoparticles led to an NADPH-dependent induction of ROS in HUVEC. ( A ) Intercellular level of NADPH analysed in lysates of control HUVEC cells (“C”) and HUVEC cells incubated for 4 h with NDs at a concentration of 20 and 50 mg/l (“ND”). Statistical significance is indicated with different superscripts: P<0.001 (a, b, c) (ANOVA; P < 0.05; n = 2 with 4 individual replicates). ( B ) Merged images of mitochondrial superoxide staining (red channel; MitoSox Red, Thermo Fisher Scientific) and transmitted light with Nomarski interference contrast of control HUVEC cells (“C”) and HUVEC cells incubated for 4 h with NDs at a concentration of 50 mg/l (“ND 50”). ( C ) Confocal images of ROS and NO in control HUVEC cells (“C”) and HUVEC cells incubated for 4 h with NDs at a concentration of 20 and 50 mg/l (“ND 20”, “ND 50”) detected using the general oxidative stress indicator CM-H2CFDA (green channel; Thermo Fisher Scientific) and the NO synthesis level indicator DAF-FM (green channel; Thermo Fisher Scientific). Cell nuclei were stained with NucRed Live 647 (red channel; Thermo Fisher Scientific). Graphs showing ROS ( D ) and NO ( E ) levels expressed as a sum of the pixel values per cell. Statistical significance is indicated with different superscripts: P<0.001 (a, b) (ANOVA; P < 0.05; n = 2 with 7 individual replicates, and each well was imaged in 4 fields of view). All values are expressed as mean ± standard deviation.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) and human mammary epithelial cells (HMEC) were obtained from Thermo Fisher Scientific, whereas HS-5 cells were obtained from ATCC (LGC Standards, Łomianki, Poland).

Techniques: Control, Incubation, Concentration Assay, Staining, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Differences in the Cell Type-Specific Toxicity of Diamond Nanoparticles to Endothelial Cells Depending on the Exposure of the Cells to Nanoparticles

doi: 10.2147/IJN.S411424

Figure Lengend Snippet: NDs increase the synthesis level of stress-related proteins. ( A ) Antibody array analysis of cytokine synthesis in control HUVEC cells (“C”) incubated with NDs at a concentration of 20 mg/l for 24 h (“ND”). One assay consists of two membranes, which are labelled “I” and “II”, and the proteins are analysed in duplicate. The localization of the positive controls: I_A1,2; I_B1,2; I_H7,8; II_A1,2; II_B1,2; II_H7,8. The localization of the selected proteins in the order of their fold (log(2)) change shown in the graph ( B ): IL-6, I_D3,4; EGF, I_F1,2; VEGF, I_G5,6; TIMP2, I_E5,6; IL-8, I_E3,4; endostatin II_H1,2; IGF-1, I_C3,4; CCL-5, I_B5,6; PECAM-1, II_F5,6; VEGF-D, I_H5,6; uPAR, II_A7,8; VEGFR3, II_C7,8; PLG, II_G1,2; MCP-1, I_G3,4; CXCL11, II_A5,6; ANGPT1, II_E1,2; TIMP1, I_D5,6; GRO A, I_A1,2; MMP-1, II_D5,6; VEGFR2, II_B7,8; TIE-2, II_G5,6; bFGF, I_H1,2; MMP-9, II_E5,6; ANGPT2 II_F1,2. The full array map is available in Tables S1 and S2 . ( C ) Images of control HUVEC tube formation (“C”) and tube formation during treatment with NDs at a final concentration of 20 mg/l (“ND”). Graphs show the mean number of junctions of HUVEC tubes in the field of view ( D ), mean tube length in the field of view ( E ), and mean number of meshes in the field of view ( F ). Data were obtained by analysing the images using ImageJ software and the Angiogenesis Analyzer macro. Statistical significance is indicated with asterisks: *P = 0.003, P = 0.002 and P < 0.001, respectively ( t -test; P < 0.05; n = 2 with 4 individual replicates). All values are expressed as mean ± standard deviation.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) and human mammary epithelial cells (HMEC) were obtained from Thermo Fisher Scientific, whereas HS-5 cells were obtained from ATCC (LGC Standards, Łomianki, Poland).

Techniques: Ab Array, Control, Incubation, Concentration Assay, Software, Standard Deviation