Fig. 2

Suppression of ERRα completely reduces the EGF treatment-induced cell proliferation of colon cancer cells. a WB for ERRα, c-Myc, cyclin D1, pERK and ERK in the HCT116 and SW480 cells treated with EGF (20/ul) at the indicated times (0.5 h, 2 h, 4 h, 6 h, and 8 h) in serum-free medium. b CCK-8 assay for the HCT116 and SW480 cells cultured with shNC or shERRα#2 (or/and 20 ng/μl EGF) for 3 d (* P< 0.05; ** P< 0.01; *** P< 0.001). The data are presented as the mean±SD of the experiments performed in triplicate. c The relative expression level of ERRα protein in shERRα#2 group was significantly lower than that in shNC group by WB assay. d Dual luciferase reporter gene assay of the SW480 cells treated with shNC or shERRα#2 (or/and 20 ng/μl EGF) in serum-free medium for 48 h. e, f Clonogenic assays and qualitative analysis of the HCT116 and SW480 cells cultured with shNC or shERRα#2 (or/and 20 ng/μl EGF) at day 7 (* P< 0.05; ** P< 0.01; *** P< 0.001). The data are presented as the mean±SD of the experiments performed in triplicate. g WB for ERRα and c-Myc in the HCT116 and SW480 cells treated with shNC or shERRα#2 (or/and 20 ng/μl EGF) in serum-free medium for 48 h. h CCK-8 assay of the HCT116 and SW480 cells treated with XCT790 (5 μM) (or/and 20 ng/μl EGF) in serum-free medium for 3 d. i Clonogenic assays of the HCT116 and SW480 cells cultured with DMSO or 5 μM XCT790 (or/and 20 ng/μl EGF) at day 7 (* P< 0.05; ** P< 0.01; *** P< 0.001). The data are presented as the mean±SD of the experiments performed in triplicate. j WB for ERRα and c-Myc in the HCT116 and SW480 cells treated with XCT790 (5 μM) (or/and 20 ng/μl EGF) in serum-free medium for 48 h