Fig. 3
The EVI1/Snail/HDAC1 complex corepressed E-cadherin expression. (a) Knockdown of EVI1 elevated E-cadherin expression but decreased N-cadherin and vimentin in 5-8F and CNE-2 cells. Overexpression of EVI1 had the opposite effects in 6-10B cells. (b) EVI1 negatively regulated E-cadherin mRNA expression. (c) EVI1 suppressed E-cadherin promoter activity in 6-10B cells. (d) The suppressive effect of EVI1 on E-cadherin expression was abolished when snail was absent. (e) EVI1 physically interacted with snail protein. (f) EVI1 downregulation-induced E-cadherin elevation was abolished when HDAC1 was overexpressed (left panel). EVI1-induced repression of E-cadherin expression was restored when HDAC1 was downregulated (right panel). (g) When EVI1 was overexpressed in 6-10B cells, the binding of HDAC1 to the E-cadherin promoter was enhanced, and acetylation of histones was decreased. (h) Suppressive activity of EVI1 in the E-cadherin promoter of sh-EVI1–5-8F cells was significantly inhibited after the knockdown of endogenous snail or/and HDAC1. (i) The existence of snail and HDAC1 was detected in immunoprecipitates obtained with antibody against EVI1 (left panel). EVI1 and snail were detected in HDAC1 immunoprecipitates (middle panel). EVI1 and HDAC1 were detected in snail immunoprecipitates (right panel)