Figure 5.

CoREST enhances the interaction of LSD1 with Snail1 and the stability of the ternary complex. (A) Snail1, WT or Tower-deleted LSD1, and CoREST were co-expressed in HEK293 cells. Cells were treated with or without MG132 for 6 h before being collected. The levels of LSD1, Snail1 and CoREST were analysed by western blotting. (B) WT or Tower-deleted LSD1 and CoREST were co-expressed in Snail1/HEK293 cells. After immunoprecipitation of LSD1, the bound Snail1 and CoREST were examined by western blotting. (C) CoREST siRNA or non-target control (NTC) was expressed in HCT116, PC3 and MDA-MB231 cells. Endogenous LSD1, CoREST and Snail1 were examined by western blotting. (D) Cells were prepared as described above in (C). After endogenous LSD1 was immunoprecipitated, the bound CoREST and Snail1 were examined by western blotting. (E) Nuclear extracts from various cancer cell lines were analysed for the levels of Snail1, LSD1 and CoREST by western blotting. Lamin A served as the nuclear marker. The expression intensity of these proteins was plotted and shown (bottom panel).