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. 2010 Jan 22;140(2):209–221. doi: 10.1016/j.cell.2009.12.040

Figure 2.

Figure 2

BRAF Inhibitors Induce CRAF Binding to BRAF

(A) WM852, D04, MM415 and MM485 cells were treated with DMSO (−), PD184352 (PD; 1 μM), sorafenib (SF; 10 μM), 885-A (1 μM) or PLX4720 (PLX; 0.3 μM) for 4 hr. Endogenous BRAF (IP: BRAF) or endogenous CRAF (IP: CRAF) were immunoprecipitated and the immunocomplexes were western blotted (WB) for BRAF or CRAF. BRAF, and CRAF levels in the cell lysates are also shown.

(B) D04 cells were treated with DMSO (−), PD184352 (PD; 1 μM), sorafenib (SF; 10 μM) and PLX4720 (PLX; 0.3μM) for 4 hr. Endogenous CRAF (IP: CRAF) was immunoprecipitated and the immunocomplexes were western blotted (WB) for BRAF or CRAF. BRAF and CRAF levels in the cell lysates are shown.

(C) SW620, HCT116 and WM1791c cells were treated with DMSO (−), PD184352 (PD; 1 μM), sorafenib (SF; 10 μM) or 885-A (1 μM) for 4 hr. Endogenous BRAF (IP: BRAF) or endogenous CRAF (IP: CRAF) were immunoprecipitated and the immunocomplexes were western blotted (WB) for BRAF or CRAF. The cell lysates were also blotted for BRAF, CRAF, phospho-ERK (ppERK) and total ERK2 (loading control).

(D) A375 cells were treated with DMSO (−), PD184352 (PD; 1 μM), sorafenib (SF; 10 μM), 885-A (1 μM) or PLX4720 (PLX; 0.3 μM) for 4 hr. CRAF (IP: CRAF) was immunoprecipitated and the immunocomplexes were western blotted (WB) for BRAF or CRAF. BRAF and CRAF levels in the cell lysates are shown.