FOXA2 and SIRT6 repress the expression of ZEB2 transcriptionally.
Notes: (A) RT-qPCR and Western blot analysis of ZEB2 expression in HepG2 cells infected with FOXA2 siRNA or SIRT6 siRNA or cotransfected with FOXA2 siRNA and SIRT6 siRNA. (B) The relative experiments were carried out in Huh7 cells. n=3 independent experiments; *P<0.05, **P<0.01. (C) The transcriptional activity of the reporter pGL3-ZEB2 plasmids was assessed in HepG2 or Huh7 cells transfected with FOXA2 siRNA or SIRT6 siRNA or cotransfected with FOXA2 siRNA and SIRT6 siRNA. The data were represented as the mean ± SD. Renilla luciferase activity was normalized to that of firefly luciferase. (D) The transcriptional activity of the reporter pGL3-ZEB2 mutant plasmids was assessed. (E) qChIP assay was performed in HepG2 or Huh7 cells to confirm the binding of FOXA2 and SIRT6 on the promoter of ZEB2. n=3 independent experiments; *P<0.05, **P<0.01. (F) The transcriptional activity of the reporter pGL3-ZEB2 plasmids was assessed in HepG2 or Huh7 cells transfected with FOXA2 plasmids or SIRT6 plasmids or cotransfected with FOXA2 plasmids and SIRT6 siRNA. (G) RT-qPCR was used to analyze MMP9 expression in HepG2 cells infected with FOXA2 siRNA or SIRT6 siRNA or cotransfected with FOXA2 siRNA and SIRT6 siRNA.
Abbreviations: HCC, hepatocellular carcinoma; qChIP, fast chromatin immunoprecipitation; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; SIRT, sirtuin; NC, negative control.