Figure 3.

Silencing of H19 affected the expression levels of EZH2/β-catenin/GSK-3β/EMT. A. RNA immunoprecipitation was performed to analyze the interaction between H19 and EZH2. Total RNA input and isotype antibody were used as positive and negative control, respectively. After immunoprecipitation, the expression of H19 was detected by qRT-PCR in input, isotype and anti-EZH2 antibody groups. The antibody against EZH2 canimmunoprecipitate H19. B. The expression of EZH2, activated β-catenin, phosphorylated GSK-3β, cyclin D1, and c-myc were significantly decreased upon silencing H19 expression, as measured by western blot. C. Knockdown of H19 suppressed N-cadherin and vimentin expression, while upregulated E-cadherin and ZO-1 measured by western blot. D. The mRNA expression level of EMT markers (E-cadherin, ZO-1, N-cadherin and vimentin) and EMT promoting transcriptional factors (Snail1, Twist1 and ZEB1) were detected by qRT-PCR after silencing of H19. Data are representative of three independent experiments and represent the mean ± SD. #P<0.05, ##P<0.01.