Fig. 2.

N(G12V)-Ras activation increases phosphorylation of JNK and ERK1,2 MAP kinases. a Conditional expression cell line STS26T-N-Ras (G12V) 15N was treated with and without doxycycline (DOX). Cells were harvested 4 and 5 days after the removal of doxycycline. Samples were analyzed by western blot and probed for the indicated protein. After each probe, the blot was stripped and reprobed for the subsequent protein. Detection of the hemagglutinin (HA) tag indicates the induction of N-Ras expression. To demonstrate activation (positive control) or inactivation (negative control) of MAP kinases, cell line TOV112D was treated (or not) with cis-Diammine-platinum (II) dichloride and samples were run in lanes 1 and 2. Data are representative of three independent experiments. b The bar graph quantitatively illustrates the means ± SEM for the three experiments described in a. Data are presented as the fold increase relative to the samples treated with doxycycline. Significant (* P value < 0.05) increases in p-ERK1,2 expression were observed on days 4 and 5 and in p-JNK1,2 expression on day 5 compared to control cells (doxycycline treated)