Figure 5. KRAS is required for both MEK/ERK and PI3K/AKT signaling in KRAS mutant NSCLC cells.

(A) Six KRAS mutant and six KRAS wild-type NSCLC cell lines were transfected with KRAS, KRAS-OTP or control siRNAs for 48 h and cell lysates were probed with the indicated antibodies. The levels of phospho-/total ERK1/2, AKT and S6 were measured for each cell line and normalized to control transfected cells. H1792 and H358 cells are displayed as exemplars of the KRAS mutant genotype. For all western blots see Supplementary Figure S9A.

(B) NSCLC cell lines were transfected with KRAS or control siRNAs for 48 h. 24 h after transfection cells were treated with either DMSO or 100 nM rapamycin. Cell lysates were probed with the indicated antibodies. The level of phospho-/total AKT was measured for each cell line and normalized to control transfected cells for each condition (+/− rapamycin). H1792 and H358 cells are displayed as exemplars of the KRAS mutant genotype. For all western blots see Supplementary Figure S9B.