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. 2020 Dec 14;10:21873. doi: 10.1038/s41598-020-78788-z

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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A discrete role for the alpha catalytic PI3K isoform in proneural (PN) GSCs. (A) Expression z-score data from TCGA (Extended Verhaak) for PIK3CA (left panel), PIK3CB (middle panel) and PIK3CD (right panel) were downloaded from the GBM-BioDP (http://gbm-biodp.nci.nih.gov/) for CL (n = 105), MES (n = 124) and PN (n = 113) subtype GBM from the HT Human Genome U133 (HT_HG-U133A) array and analysed using GraphPad Prism 8. As recent evidence suggests the neural subtype represents excessive contamination with normal brain, it has been excluded from the analysis. Unpaired one-way ANOVA, *p ≤ 0.05; **p ≤ 0.01; ****p ≤ 0.0001. (B) Survival analysis based on the impact of the multi-gene prognostic index for coexpression of PIK3CA and NES (upper panels), PIK3CB and NES (middle panels) or PIK3CD and NES (lower panels) for CL (n = 53, left panels), MES (n = 57, middle panels) and PN (n = 56, right panels) subtype. TCGA gene expression data (Verhaak Core) for PIK3CA and NES, PIK3CB and NES or PIK3CD and NES from the HT_HG-U133A array were used for multigene prognostic index. Figure was generated using the GBM-BioDP software. (C) GSC lines JK16, 83Mes, 157PN and AC17PN were analysed by Western blot using antibodies against NF1, p110α and GAPDH. Membrane was stripped and reprobed with an antibody against p110β. Lysates from the same experiment were run in parallel and probed for p110 $δ$ and GAPDH. (D) Reverse-phase protein microarray (RPPA) data from TCGA GBM samples. Boxplots of RPPA analytes for phospho-AKT(Ser⁴⁷³) measured in CL (n = 28), MES (n = 29) and PN (n = 41) samples from the cohort of GBM tumours (Extended Verhaak) that was subjected to RPPA analysis within the TCGA consortium. RPPA data were downloaded from the GBM-BioDP and analysed using GraphPad Prism 8. Unpaired one-way ANOVA: *p ≤ 0.05. (E) Survival analysis based on RPPA analysis for phospho-AKT(Ser⁴⁷³) using the Extended Verhaak cohort submitted to the TCGA consortium for CL (n = 28, left panel), MES (n = 29, middle panel) and PN (n = 41, right panel) subtypes. Figure was generated using the GBM-BioDP software. (F) GSC lines JK16, 83Mes, 157PN and AC17PN were analysed by Western blot using antibodies against AKT. Membrane was stripped and re-probed for p-AKT(Thr³⁰⁸), then stripped and reprobed for p-AKT(Ser⁴⁷³), then stripped and re-probed for HSP90.