Extended Data Fig. 10: Biological relevance of PIK3CA MMs.
From: Landscape and function of multiple mutations within individual oncogenes
a, Viability of Ba/F3 cells expressing mock control or wild-type, single-mutant or double-mutant PIK3CA without WEHI-3-conditioned medium for three days (n = 3). Data represent means + s.d. Two-sided Welch’s t-test. b, Dependency of 196 CCLE cell lines on AKT1, with cell lines coloured by PIK3CA MM status, in the DepMap CRISPR–Cas9 knockout data (see Methods). Five cancer types for which there are cell lines harbouring PIK3CA MMs were analysed by two-sided Mann–Whitney U-test. c, Box plots showing sensitivity to six PI3K inhibitors (Z-scores) for 417 CCLE cell lines, according to MM status. Five cancer types for which there are cell lines harbouring PIK3CA MMs were analysed. d, Immunoblot analysis of AKT, pS473 AKT, PIK3CA and β-actin in Ba/F3 cells expressing mock control or wild-type, single-mutant or double-mutant PIK3CA in the presence of WEHI-3-conditioned medium. Representative of three independent experiments. e, Capillary-based immunoassay of AKT, pS473 AKT, mTOR, pS2448 mTOR, p70 S6K, pT389 p70 S6K, PRAS40, pT246 PRAS40, GSK-3β, pS9 GSK-3β, PIK3CA and β-actin in Ba/F3 cells expressing mock control or wild-type, single-mutant or double-mutant PIK3CA without WEHI-3-conditioned medium. Representative of two independent experiments. d, e, See Supplementary Fig. 1 for source images. f, g, Box plots showing levels of phosphorylated S473 (pS473; left) and T308 (pT308; right) in RPPA data relative to total AKT protein expression for 316 CCLE cell lines (with cancer types for which there are cell lines harbouring PIK3CA MMs) (f) and in 3,164 TCGA patients (with purity of 50% or more, in recurrently mutated cancer types) (g) according to MM status. Each dot represents a sample, coloured by cancer type. c, f, g, Single mutations were classified into missense mutations in major and minor hotspot positions, in-frame indels, and non-hotspot/functional mutations, some of which are not shown owing to small numbers. Box plots show medians (lines), interquartile ranges (IQRs; boxes) and 1.5 × IQRs (whiskers). GLM, generalized linear model (see Methods). Examined numbers are shown in parentheses.
