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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 2. Proliferation of M14 AZ628-resistant (M14BRR) clones is dependent on MEK but not BRAF.

Figure 2

(A) Dose-response curves of M14 and three M14BRR clones treated with the indicated concentrations of the MEK inhibitor U0126. The percentage of viable cells is expressed relative to untreated controls. Error bars represent the standard deviation from the mean. The A431 cell line survival curve is shown as a negative control.

(B) AZ628-4resistant cells retain sensitivity to U0126. Cell lysates from M14, AZ628-resistant clones, and A431 (negative control) were collected following treatment with the indicated concentrations of AZ628 or U0126 for 2 hours. Immunobloting analysis was performed using antibodies directed against the indicated proteins.

(C) Effective depletion of BRAF protein by shRNA. M14 and M14BRR2 cells were infected with lentivirus containing control (PLKO.1 empty vector) or BRAF-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.

(D) Reduced dependency on BRAF in AZ628-resistant cells. Proliferation assay corresponding to cells in (C). Control or BRAF-specific BRAF shRNAs were introduced in A549, M14, and M14BRR2 cells by lentiviral infection and a cell proliferation assay with Syto60 was performed 7 days later. The fraction of viable cells is expressed relative to untreated control. Error bars represent the standard deviation from the mean.