Fig. 6
YAP bound and sustained HIF-1α stability to promote glycolysis in HCC cells under hypoxia. a, b Representative immunostaining of YAP and HIF-1α in HepG2 and Huh7 cells under normoxia (20%O2) or hypoxia (1%O2) for 24 h. (magnification: × 200). c Co-immunoprecipitation of endogenous YAP and HIF-1α in HCC cells under hypoxia (1%O2) for 24 h in the presence of 10 μM MG132. d Western blot showed the HIF-1α stability in control or siYAP cells treated with 100 μg/ml cycloheximide (CHX) at 0, 2 h and 4 h. e ChIP assay was performed with antibody against HIF-1α, YAP or control IgG in Huh7 cells under normoxia (20%O2) and hypoxia (1%O2) for 24 h. Real-time PCR was used to analyze the immunoprecipitated DNA. *p < 0.05 versus IgG. f ChIP assay was performed with antibody against HIF-1α, YAP or control IgG in Huh7 cells under hypoxia (1%O2) for 24 h after transfected with YAP siRNA. Real-time PCR was used to analyze the immunoprecipitated DNA. ns > 0.05, *p < 0.05 versus IgG. g pGL2p control (con), pGL2p-WT PKM2 HRE (wt), or pGL2p mutant PKM2 HRE (mut) with CGT/AAA were cotransfected into Huh7 cells under normoxia (20%O2) or hypoxia (1%O2) for 24 h. Dual-luciferase reporter assay was performed to detect the promoter activity, which was normalized to Renilla luciferase activity. *p < 0.05, **p < 0.01 h A diagram about the potential mechanism. All data are shown as the mean ± SEM of three independent experiments