(A) CRAF cDNA or control (pBABE empty vector) were introduced into M14 parental cells. Cell lysates were collected and immunobloting analysis was performed using antibodies directed against CRAF and t-ERK 1/2 (loading control).
(B) M14 cells expressing exogenous CRAF exhibit reduced sensitivity to AZ628. Dose-response curves corresponding to cells in (D). M14+CRAF and M14+pBABE control vector cells were treated with the indicated concentrations of AZ628. The fraction of viable cells is expressed relative to untreated controls. Error bars represent the standard deviation from the mean.
(C) M14 cells expressing exogenous CRAF exhibit reduced suppression of ERK1,2 activation following AZ628 treatment. Cell lysates from M14+pBABE control vector and M14+CRAF were collected following treatment with the indicated concentrations of AZ628 for 2 hours. Immunobloting analysis was performed using antibodies directed against the indicated proteins.