BRAF Binding to CRAF Requires BRAF and CRAF Binding to RAS, Relates to Figure 3
(A) Myc-epitope tagged CRAF or R89LCRAF (R89L), or an empty vector control (EV) were transfected into D04 cells. After 48 hr the cells were treated with DMSO (-) or sorafenib (SF, +; 10 μM) for 4 hr. The myc-CRAF was immunoprecipitated (IP) and the immunocomplexes were western blotted (WB) for endogenous BRAF or myc-CRAF. Endogenous BRAF and myc-CRAF levels in the cell lysates are also shown.
(B) Myc-epitope tagged BRAF or R188LBRAF (R188L) or an empty vector control (EV) were transfected into D04 cells. After 48 hr the cells were treated with DMSO (-) or sorafenib (SF, +; 10 μM) for 4 hr. The myc-BRAF was immunoprecipitated (IP) and the immunocomplexes were western blotted (WB) for myc-BRAF or endogenous CRAF. Myc-BRAF and endogenous-CRAF levels in the cell lysates are also shown.