Fig. 4

Ezh2 regulates PTEN/AKT signaling by directly binding to the promoter regions of PTEN in GC. a Representative images of the Western blot analysis for expression of Ezh2, PTEN, p-Akt, and total Akt in Ezh2-overexpressing MKN-45 and SGC-7901 cells and normal control, as well as Ezh2-knockdown AGS cells and normal control. b Representative images of the Western blot analysis for basic expression of Ezh2 and PTEN in five GC cell lines and the normal human gastric mucous cell line (GES-1). c Representative images of the IHC analysis for expression of Ezh2, PTEN, p-Akt, and total Akt in xenograft tissues. d Ezh2 and PTEN mRNA expression correlation analyses using the TCGA gastric cancer data. e The qRT-PCR results showed that PTEN mRNA was decreased in Ezh2-overexpressing MKN-45 and SGC-7901 cells, while increased in Ezh2-knockdown AGS cells. Data are represented as mean ± SEM. *P < 0.01. f Dual-reporter luciferase assays showed that overexpression of Ezh2 in HEK-293T and MKN-45 cells suppressed the promoter activity of PTEN. Data are represented as mean ± SEM. *P < 0.05. g Represent schemata of the PTEN promoter regions with or without binding affinity for EZH2. Arrow indicates the transcriptional start site. ATG indicates translation start codon. h ChIP assays showed that endogenous Ezh2 bound to the promoter region of PTEN. IgG served as a negative control, and H3K27 (H3) served as a positive control