(A) Pik3caWT/Flox and Pik3caMUT/Flox MEFs (expressing ROSA26 Cre-ER) were treated overnight with 1 μM tamoxifen or vehicle to generate Pik3caWT/− and Pik3caMUT/− MEFs. Cells were starved overnight and stimulated for 10 min with epidermal growth factor (EGF) (10 ng/mL) or vehicle. Proteins were collected and subjected to western blot analysis as indicated. One representative experiment from three is shown.

(B and E) PC9 (B) and H1975 (E) EGFR-mutant lung cancer cells were infected with retroviruses encoding Pik3caWT or Pik3caMUT or not infected (parental). Cells were then starved and stimulated with EGF (10 ng/mL) or vehicle for 10 min. Proteins were collected and subjected to western blot analysis as indicated. One representative experiment from three is shown.

(C) Quantification of the western blots in (B) (n = 3).

(F) Quantification of the western blots in (E) (n = 3).

(D and G) Cell growth assay by crystal violet staining in PC9 (D) and H1975 (G) cells expressing Pik3caWT, Pik3caMUT or none (parental). Means of three replicates are shown.