Fig. 6

The lncRNA-p21 competes with Hotair to interact with EZH2 and promotes phosphorylation of EZH2 at S21. a, b RNA-IP was performed to detect the lncRNA-p21 interaction with EZH2 by using lncRNA-p21 anti-sense probe vs sense probe in (a) C4-2 cells and in (b) Enz-treated C4-2 cells. c RNA in situ hybridization and immunofluorescent staining to detect the co-localization of lncRNA-p21 and EZH2 in C4-2 cells w/o Enz treatment (Scale bar = 2 μm). d In C4-2 cells, the EZH2 was pulled down by EZH2 antibody and the levels of lncRNA-p21 and Hotair were detected by qPCR. e, f RNA-IP assay to detect the interaction between Hotair and EZH2 after treating w/o Enz (e) and w/o overexpression of lncRNA-p21. f, g STAT3 methylation levels were detected by WB in C4-2 pLKO and sh-EED cells after 4 days w/o Enz treatment. h EZH2 p-S21 levels were detected in C4-2 PWPI and PWPI-lncRNA-p21 cells. i EZH2 p-S21 level was detected in C4-2 pLKO and sh-lncRNA-p21 cells w/o Enz treatment. j The Co-IP assay to detect the AKT and EZH2 interaction in C4-2 pWPI and C4-2 pWPI-lncRNA-p21 cells. k C4-2 pLKO and sh-lncRNA-p21 cells were treated w/o Enz for 4 days and then EZH2 was immunoprecipitated. The AKT and EZH2 interaction was analyzed by WB. l C4-2 cells were treated w/o Enz, and then the lncRNA-p21 was pulled down. The interaction of AKT and EZH2 with lncRNA-p21 was analyzed by WB. For d, the data are presented as mean ± SD, *p < 0.05, **p < 0.005, by ANOVA