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. Author manuscript; available in PMC: 2011 Sep 22.
Published in final edited form as: Nature. 2010 Mar 18;464(7287):427–430. doi: 10.1038/nature08902

Figure 2. MEK/ERK activation requires binding of drug to the catalytic domain of RAF.

Figure 2

a, 293H cells transfected with EGFP (control), HA-tagged RASG12V, the catalytic domain of CRAF (V5-tagged catC) and catC carrying a mutation at the gatekeeper residue (V5-tagged catCT421M), treated with vehicle or PLX4720 (1μM/1 hour). Lysates were subjected to immunoblot analysis for pMEK and pERK. b, Wild-type (+/+), BRAF knock-out (BRAF −/−) or CRAF knock-out (CRAF −/−) mouse embryonic fibroblasts (MEFs) were treated with the indicated concentrations of PLX4720 for 1 hour. c, Sorafenib inhibits the gatekeeper mutant catCT421M protein in vitro (Supplementary Fig. 8c) and activates MEK/ERK in cells expressing it. 293H cells overexpressing catCT421M were treated with the indicated concentrations of sorafenib for 1 hour. Lysates were subjected to analysis for pMEK and pERK.