Fig. 3: PTEN loss activates AKT but not mTORC1 in ASP^cKO tumors.

a Heatmap of the 55 significantly (P < 0.05) different total and phosphoproteins by RPPA, normalized, log2 transformed and median-centered between ASP^WT and ASP^cKO tumors, n = 3. b Immunoblots of PTEN, phosphorylated AKT^Ser473 or AKT^Thr308, AKT, phosphorylated S6^Ser235/236, S6, phosphorylated-4E-BP1, 4E-BP1, phosphorylated p70S6K, p70S6K, and GAPDH, n = 3. c–f CellTiterGlo growth assays of ASP^WT or ASP^cKO rhabdospheres treated with GDC-0941 (c) (P = 0.0020), MK-2206 (d), everolimus (e), or doxorubicin (f), n = 3, in triplicate. g Principal component analysis of transcriptional profiles from ASP^WT (black) and ASP^cKO (blue) tumors and Cre-negative SCM (green), n = 4. h Volcano plot showing differentially expressed genes between ASP^WT and ASP^cKO tumors from (g). Genes with increased (red) and decreased (blue) expression in ASP^cKO tumors (≥twofold, P < 0.05) are highlighted. i Validation of gene expression in (h) by real-time PCR (n = 3 SCM, n = 4 tumors, in duplicate), expression normalized to 18S rRNA and expressed relative to ASP^WT. (Dlx5 P = 0.0062, Mmp9 P = 0.0009, Foxm1 P = 0.0011, Bub1b P = 0.0002, Ccnb1 P = 0.0002, Tyms P = 0.0009, Pvalb P = 0.0001, Apod P = 0.0018). j, k Gene ontology (GO) analyses of genes with increased (j) and decreased (k) expression in ASP^cKO tumors versus ASP^WT tumors. Top ten unique GO terms are shown. All P values in pairwise comparisons were determined by Student’s t test (unpaired, two-tailed); **P < 0.01, ***P < 0.001, ****P < 0.0001. Data represented as mean + SEM. See also Supplementary Figs. 4 and 5 and Supplementary Data 1. Source data are provided as a Source Data file.