Journal: Scientific Reports
Article Title: Combined PI3Kα-mTOR Targeting of Glioma Stem Cells
doi: 10.1038/s41598-020-78788-z
Figure Lengend Snippet: 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 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\updelta $$\end{document} δ and GAPDH. ( D ) Reverse-phase protein microarray (RPPA) data from TCGA GBM samples. Boxplots of RPPA analytes for phospho-AKT(Ser 473 ) 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(Ser 473 ) 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(Thr 308 ), then stripped and reprobed for p-AKT(Ser 473 ), then stripped and re-probed for HSP90.
Article Snippet: Similarly, protein Reverse Phase Protein Microarray (RPPA) data of the Extended Verhaak dataset for p-AKT (S473) were downloaded and analysed using GraphPad Prism 8.
Techniques: Expressing, Gene Expression, Generated, Software, Western Blot, Membrane, Microarray
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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 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\updelta $$\end{document} δ and GAPDH. ( D ) Reverse-phase protein <t>microarray</t> <t>(RPPA)</t> data from TCGA GBM samples. Boxplots of RPPA analytes for phospho-AKT(Ser 473 ) 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(Ser 473 ) 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(Thr 308 ), then stripped and reprobed for p-AKT(Ser 473 ), then stripped and re-probed for HSP90.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6588/pmc07736588/pmc07736588__41598_2020_78788_Fig4_HTML.jpg)