Fig. 5
From: PDCD6 cooperates with C-Raf to facilitate colorectal cancer progression via Raf/MEK/ERK activation
PDCD6 is physically associated with c-Raf and activates the MAPK/ERK signaling pathways in CRC cells. a. Coimmunoprecipitation assays using anti-PDCD6 antibody and anti-c-Raf antibodies showed that PDCD6 interacts with c-Raf in HCT116 cells. b. Confocal IF staining of PDCD6(green) and c-Raf (red) in HCT116 cells. The nuclei were stained by DAPI (blue). The results from one of two comparable experiments are shown. c. HCT116 cell lysate was immunoprecipitated with anti-PDCD6 and anti-c-Raf antibodies in the presence, and absence of EDTA. d. Immunoblot analysis of the expression of PDCD6 and the total and phosphorylated levels of c-Raf/MEK/ERK in HCT116 cells with PDCD6 knocked down and overexpression. Tubulin was used as a loading control. Bands were quantified with ImageJ software. e. Immunoblot analysis of PDCD6 the total and phosphorylated level of c-Raf/MEK/ERK in HCT116 cells overexpressing PDCD6 in the presence of the Ca2+ chelator BAPTM. f. Immunoblot analysis of PDCD6, total MEK/ERK and phosphorylated MEK/ERK in cells with PDCD6-OE, PDCD6-OE + RAF709 (R) and PDCD6-OE + Trametinib (Tr), respectively. Tubulin was used as a loading control