Fig 1: E-cadherin interacts with AGO2 and upregulates its protein expression. Effect of E-cadherin on protein expression of cytoplasmic miRNA biogenesis factors (A-D). (A) AGO2, GW182, Dicer, TRBP, FMRP, SND1 and PACT protein expression were determined by western blot in E-cadherin-overexpressing HeLa cells. (B) AGO2 were determined by western blot in HeLa cells with sequential increase of E-cadherin overexpression. (C) Effects of AGO2 knockdown on protein expression of AGO2, GW182, Dicer and TRBP in E-cadherin-overexpressing HeLa cells. AGO2 was further knocked down by specific shRNA in E-cadherin-overexpressing HeLa cells. (D) E-cadherin was knocked down in MCF-7 cells using specific shRNAs. Protein expression of AGO2, GW182, Dicer and TRBP were determined by western blot. Effects of E-cadherin on mRNA expression of AGO2 (E,F). (E) AGO2 mRNA expression was determined by real-time quantitative reverse-transcription PCR (qRT-PCR) in E-cadherin-overexpressing HeLa cells. (F) E-cadherin was knocked down in MCF-7 cells using specific shRNAs. mRNA expression of AGO2 was determined by qRT-PCR. Data were at least repeated in three independent experiments (mean ± SD) and statistically analyzed by two-tailed Student’s t-test (E) and one-way ANOVA (F). (G) Interaction between E-cadherin and AGO2 in E-cadherin-overexpressing HeLa cells. Immunoprecipitation were performed using anti-E-cadherin antibody. (H) Effects of AGO2 knockdown on the interaction between E-cadherin and AGO2. AGO2 was knocked down by specific shRNA in E-cadherin-overexpressing HeLa cells. Immunoprecipitation was performed using anti-E-cadherin antibody. (I) Effects of PLEKHA7 knockdown on the interaction between E-cadherin and AGO2. PLEKHA7 was knocked down by specific shRNA in E-cadherin-overexpressing HeLa cells. Immunoprecipitation was performed using anti-E-cadherin antibody.