Figure 4: Switch 2 insertion mutations alter PI3K interaction and result in altered effector signalling dependencies.
From: KRAS insertion mutations are oncogenic and exhibit distinct functional properties
(a) Western blot (top) and quantification (bottom) of lysates prepared from Ba/F3 cells expressing wild-type K-Ras (WT), K-RasG12D (D12), K-RasQ61L (L61), K-RasE62_A66dup (5AA), or K-RasG60_A66dup (7AA) after a 6 h starve in media containing 0.5 μg ml−1 doxycycline, no IL-3 and 1% FBS. Quantification shows the average±s.e.m. of four independent experiments. (b) Representative western blots (top) and quantifications (bottom) of recombinant effector binding to His–K-Ras proteins loaded with the indicated nucleotide. Quantification shows the average±s.d. of two independent experiments. (c,d) Dose response curves of Ba/F3 cells transformed with various K-Ras proteins that were grown for 3 days in the presence of the indicated concentrations of either PD0325901 (PD-901) (c) or GDC-0941 (d). Data are mean±s.e.m. of three wells and are representative of three independent experiments. Trend lines represent best fit to a fixed slope sigmoidal dose response equation. The 95% CI of GI50 values are indicated. a.u., arbitrary unit; CI, confidence interval.
