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. 2005 Dec 21;103(1):57–62. doi: 10.1073/pnas.0609973103

Fig. 4.

Fig. 4.

V600E B-Raf mutation confers sensitivity to 17-AAG-induced inhibition of kinase activity and protein expression. Myc-tagged WT or V600E mutant B-Raf was transiently expressed in MCF-7 cells. At 24 h after transfection, cells were treated with 1 μM 17-AAG for 12 or 24 h. (A) Expression of myc-tagged WT and V600E mutant B-Raf was analyzed by immunoblotting (Top). The effect of 17-AAG on endogenously expressed WT B-Raf (Middle) and C-Raf (Bottom) were also assessed. (B) Results of a representative experiment were quantified and are presented graphically as the relative change in expression levels of indicated proteins vs. control. (C) Kinase activity of B-Raf assayed by IP with myc after 17-AGG treatment over 24 h was measured by using a kinase assay.