A, Cell Titer-Glo cell viability assay after 6-day temozolomide treatment for glioma tumorsphere lines. *, ** P<0.05 for the difference between DMSO and TMZ at indicated concentration in MGG152 (*) and YMG12 (**). B, Complete remission of recurrent AOD after 4-cycle (8 months) of a chemotherapeutic regimen consisting of procarbazine, nimustine (ACNU), and vincristine. Contrast enhanced MRI showing rapid recurrence (yellow circles) 2 months after gross total resection (GTR) of YMG6R4T (left) and post-chemotherapy (right). C, Cell Titer-Glo cell viability assay after 3-day treatment of anaplastic oligodendroglioma lines (YMG6R4T and YMG23, both PIK3CA mutant) and a glioblastoma line (YMG36) with ACNU, CCNU, procarbazine, or vincristine. *, **, *** P<0.05 for the difference between DMSO and indicated chemotherapeutic agent in YMG6R4T (*), YMG23 (**), and YMG36 (***). D, Relative cell viability of YMG6R4T (left) and YMG23 (right) cells after 3-day treatment with FK866 combined with DMSO control (blue bars) or TMZ (200 μM, purple bars). *, P<0.05 for difference DMSO and FK866. **, P<0.05 for difference DMSO and FK866 plus TMZ. E, Cell viability of YMG6R3T, YMG6R4T and YMG23 cells (all PIK3CA mutant) after 9-day exposure with AGI-5198 (IDH1R132H specific inhibitor), relative to DMSO control. F, Western blot analysis of p-AKT, p-mTOR, p-4EBP1, and p-S6K expression in YMG6R4T cells after 12 hr-treatment with DMSO control, LY294002 (PI3K inhibitor, 50 μM, left), and GDC-0068 (AKT inhibitor, 5 μM, right). G, Cell Titer-Glo cell viability assay after 3-day treatment of YMG6R4T, YMG23, and YMG28 (PI3K pathway gene wild-type) AOD cells with LY294002 or GDC-0068. *, ** P<0.05 for the difference between DMSO and treatment at indicated concentration in YMG6R4T(*) and YMG23 (**). H, Cell Titer-Glo cell viability assay after 3-day treatment of YMG6R4T, YMG23, and YMG28 (PI3K pathway gene wild-type) AOD cells with BYL719 (PI3K inhibitor) or everolimus (mTOR inhibitor). *, ** P<0.05 for the difference between DMSO and treatment at indicated concentration in YMG6R4T(*) and YMG23 (**). I, Cell Titer-Glo cell viability assay after 3-day treatment of YMG46 (PIK3CAGlu469Gly, not reported in COSMIC database) AOD cells with LY294002, GDC-0068, BYL719, or everolimus. * P<0.05 for the difference between DMSO and treatment at indicated concentration. J, Kaplan-Meier curves indicating survival difference between mice implanted with DMSO (24hr, n=3) or LY294002 (50 μM, 24hr, n=3) pretreated YMG6R4Tsc2 cells (2×105 cells/mouse). Bars, SEM.