Figure 6. FOXA2 is required for demethylation-mediated E-cadherin re-activation.
(A, B) 5′-Aza-dC treatment induces E-cadherin and FOXA2 expression. MiaPaCa-2 cells were treated with 5′-Aza-dC for 5 days at indicated doses. Expression of E-cadherin, FOXA1 and FOXA2 were examined by qRT-PCR (A) and flow cytometry (B). (C, D) 5′-Aza-dC treatment induced E-cadherin expression is dependent on FOXA2 elevation. MiaPaCa-2 cells were treated with 1 μM 5′-Aza-dC for 5 days. On the 3rd day, two different synthetic siRNAs (#7 and #8) against FOXA2 were independently transfected into 5′-Aza-dC treated or untreated MiaPaCa-2 cells. Expression of E-cadherin and FOXA2 were examined by qRT-PCR (C) and flow cytometric analysis (D). For qRT-PCR analysis, mean±SEM from three independent experiments are shown. For FACS analysis, representative data from at least three independent experiments are shown. Only four data lines are shown for simplicity (representative complete data set is presented in Fig S4). Bar graph summarizes at least three independent experiments. *: p<0.05; **: p<0.01.