Fig. 6

DRAM1 inhibits serum-stimulated phosphorylation of rpS6 and reduces cell proliferation under serum starvation condition. a HEK293T cells were transfected with FLAG empty vector or FLAG-DRAM1 for 24 h followed by starvation for another 18 h before stimulating with serum (10%) over a time-course. Both phosphorylation sites of rpS6 (S235/236 and S240/244) were detected with phospho-specific antibodies. Total protein levels of rpS6, FLAG and β-actin were also detected with immunoblotting. b and c HEK293T and SW480 cells were transfected with FLAG empty vector or FLAG-DRAM1 plasmids for 24 h, and the cells were starved for another 24 h before examining cell viability using CCK-8 kit. d and e HEK293T cells were transfected with FLAG vector or FLAG-DRAM1 for 24 h, and serum starved for another 24 h, then the cells were counted and seeded onto 6-well plate. Two weeks late, the colony number was counted. *p < 0.05 and **p < 0.01 vs the indicated groups