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. Author manuscript; available in PMC: 2009 Jun 8.
Published in final edited form as: Cancer Res. 2008 Jun 15;68(12):4853–4861. doi: 10.1158/0008-5472.CAN-07-6787

Figure 4. Proliferation of AZ628-resistant M14 clones is dependent on CRAF.

Figure 4

(A) Down-regulation of CRAF in AZ628-resistant clones results in reduced p-ERK1/2. M14 and M14BRR2 cells were infected with a lentivirus control (PLKO.1 empty vector) or a virus expressing CRAF-specific shRNA. Cells were puromycin-selected and protein lysates were collected 4 days after the infection. Immunobloting analysis was performed using antibodies directed against the indicated proteins.

(B) AZ628-resistant M14 cells are dependent on CRAF. Cell viability assay corresponding to (A). Control or CRAF-specific shRNAs were introduced into A549, M14, M14BRR2, and M14BRR8 cells and cell proliferation assays with Syto60 were performed 5 days later. The fraction of cells relative to untreated controls is expressed. Error bars represent the standard deviation from the mean.

(C) CRAF levels in AZ628-resistant cells vary proportionately to the concentration of AZ628 in which cells are maintained. Cell lysates from M14BRR2 cells growing in the indicated concentrations of AZ628 for several passages were collected. Immunobloting analysis was performed using antibodies directed against the indicated proteins.