Fig. 1

D594A mutation confers lower aggressiveness on MC38 cells. A Immunoblotting of BRAF level in WT and mutant MC38 cells. Actin serves as an internal reference. B Representative immunoblotting of MEK, pMEK, ERK1/2, pERK1/2 and Cyclin D1 in the starvation group (right) and stimulation group (left). Cells were exposed to EGF (100 ng/mL) for 15 min after starvation for 36 h in stimulation group. C Proliferation ability of BRAF WT and mutant MC38 cells at 24 h, 48 h and 72 h detected by CCK-8 assay. D Detection of apoptosis by annexin V/PI staining after induction of FCCP for 36 h. E Growth ability of BRAF WT and mutant MC38 cells analyzed by clone formation assay. F, G Migration and invasion abilities of BRAF WT and mutant MC38 cells analyzed by transwell assays, respectively. Three independent replicates were performed for above experiments. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001