clonogenic cell survival assay cell viability  (Dojindo Labs)

Journal: Hepatology (Baltimore, Md.)

Article Title: HMGB1 Controls Liver Cancer Initiation through YAP-dependent Aerobic Glycolysis

doi: 10.1002/hep.29663

Figure Lengend Snippet: (A) Q-PCR analysis of mRNA expressions of indicated genes in Hepa1-6 and HuH7 cells after knockdown of HMGB1 (n=3, *p<0.05 versus control shRNA group). (B, C) Western blot analysis of indicated protein expression in Hepa1-6 and HuH7 cells after knockdown of HMGB1 or YAP. (D) Forced expression of HMGB1 cDNA restored HMGB1 and YAP expression in HMGB1−/− MEFs. (E) Forced expression of YAP cDNA restored YAP expression, but not HMGB1 expression in indicated HMGB1-knockdown HCC cells. (F) Forced expression of YAP reversed cell proliferation inhibition in HMGB1 knockdown HCC cells (n=3, *p < 0.05). (G–H) Clonogenic cell survival assay determines the reproductive ability of indicated HCC cells. A representative image is shown in panel G. Relative reproductive ability is semi-quantified in panel H (n=3, *p < 0.05).

Article Snippet: Cell viability and clonogenic cell survival assay Cell viability was assayed using Cell Counting Kit-8 kits (Dojindo Laboratories).

Techniques: Knockdown, Control, shRNA, Western Blot, Expressing, Inhibition, Clonogenic Cell Survival Assay

Journal: Hepatology (Baltimore, Md.)

Article Title: HMGB1 Controls Liver Cancer Initiation through YAP-dependent Aerobic Glycolysis

doi: 10.1002/hep.29663

Figure Lengend Snippet: (A) Indicated mouse and human HCC cells were treated with glycyrrhizin (0.625, 1.25, 2.5, 5, 10 mM) or verteporfin (2.5, 5, 10, 20, and 40 µM) for 24 hours. Cell viability was assayed (n=3, *p<0.05 versus control untreated group). (B) Clonogenic cell survival assay determined the reproductive ability of Hepa1-6 and HuH7 cells following treatment with glycyrrhizin (2.5 mM) or verteporfin (10 µM). A representative image is shown in the left panel. Relative reproductive ability is semi-quantified in the right panel (n=3, *p < 0.05 versus control untreated group). (C) Q-PCR analysis of mRNA expressions of indicated genes in HCC cells after glycyrrhizin (2.5 mM) or verteporfin (10 µM) treatment for 24 hours (n=3, *p<0.05). (D–F) ECAR levels in indicated HCC cells (n=3, *p < 0.05). (G) HIF1α DNA binding activity in indicated HCC cells with or without glycyrrhizin (2.5 mM) or verteporfin (10 µM) treatment for 24 hours (n=3, *p<0.05 versus control in the treated group). (H) Immunoprecipitation analysis of HIF1α-YAP complex in hepatocytes from DEN-induced Hmgb1f/f mice or primary human HCC (pHCC) cells or HuH7 cells. (I–K) YAP-knockdown HuH7 cells were transfected with YAP-cDNA or YAP-5SA-S94A mutant for 48 hours, and then treated with hypoxia (1% O2) for 24 hours. HIF1α DNA binding activity (I), gene expression (J), and YAP-HIF1α complex (K) were assayed.

Article Snippet: Cell viability and clonogenic cell survival assay Cell viability was assayed using Cell Counting Kit-8 kits (Dojindo Laboratories).

Techniques: Control, Clonogenic Cell Survival Assay, Binding Assay, Activity Assay, Immunoprecipitation, Knockdown, Transfection, Mutagenesis, Gene Expression