Fig. 3

K-Ras(V12) differentially regulates Akt isoform expression. The amount of Akt isoforms was analyzed on protein (A, B) and mRNA (C, D) level in PANC-1 and Colo-699 cancer cells. A, B Akt isoforms were detected in RIPA cell lysates by Western blotting using Akt isoform-specific antibodies and GAPDH as loading control. Depicted are representative blots of at least three independent experiments. The bar graphs show the densitometric analysis as a ratio of Akt in relation to loading control normalized to an EGFP control clone 2 (mean ± SD, n = 4–5). C, D The mRNA content of each AKT variant was determined by qRT-PCR analysis. Relative expression of each AKT variant was assessed in relation to the expression of the house keeping gene RPLP0 and normalized to the corresponding EGFP cell clone. Each sample was analyzed in duplicate using three (Colo-699) or 3–6 (PANC-1 clones) different cDNAs (mean ± SD). Statistical analyses: one sample t-test, **** p ≤ 0.0001; *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05