Fig. 4

PTENp1 expression is regulated by TETs. a Hepatocytes and HCC cells were subjected to methylation-specific PCR assay. b TET1, TET2 and TET3 expression was measured by real-time qPCR analysis. c-e Hep3B cells were transfected with TET1-O (TET1 overexpression vector), TET2-O, TET3-O, TET1 siRNA, TET2 siRNA or TET3 siRNA and then TET1, TET2 and TET3 expression was measured by western blot analysis. f Hep3B cells were transfected as indicated, and then PTEN and PTENp1 expression was measured by real-time qPCR analysis. g PTEN expression was measured by western blot analysis. h Hep3B cells were transfected with TET1-O + PTENp1 siRNA, TET2-O + PTENp1 siRNA, TET3-O + PTENp1 siRNA, and PTENp1 siRNA alone and then PTEN and PTENp1 expression was measured by real-time qPCR analysis. i Hep3B cells were transfected as an indication, and then PTEN expression was measured by western blot analysis. j Hep3B cells stably infected with TET1, TET2, TET3 or TET1 shRNA lentivirus were subjected to methylation-specific PCR assay. k ChIP assay using anti-TET1, anti-TET2, anti-TET3 or IgG in Hep3B cells and qPCR of the PTENp1 promoter. Values correspond to the ratio between the anti-TET1, anti-TET2 or anti-TET3 antibody immunoprecipitated DNA relative to the IgG immunoprecipitated DNA. The values expressed are the mean ± SD from three samples (^*P < 0.01, vs. normal, control or IgG)