Fig 1: Screening of downstream targets of CD97 in GSCs(A) Schematic representation of the experimental strategy for the transcriptome analysis. Heatmap of the hierarchical clustering analysis showing the expression of 131 common downregulated genes in CD97-knockdown GSCs (right).(B) Gene set enrichment analysis (GSEA) plot for the regulation of stem cell population maintenance signature from the Gene Ontology Biological Process (GOBP) category comparing the shCtrl and shCD97 groups. NES, normalized enrichment score.(C) Bubble chart showing the top 10 biological processes of Gene Ontology terms for genes downregulated in CD97-knockdown GSCs. Size, gene number; color, −log10 (p value).(D) RT-qPCR of PCBP1, BSG, ARHGAP1, BZW1, BZW2, and DBN1 expression in three different GSCs (83, X01, and 528 cells) infected with shCtrl or shCD97 lentivirus. β-actin was used as a loading control. All error bars represent mean ± SD (n = 3 independent experiments). ∗∗∗∗p < 0.0001, ∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05, t test.(E) Kaplan-Meier overall survival curves for patients with GBM presenting the high and low expression of the CD97/ARHGAP1 (left), CD97/BZW1 (middle), and CD97/BZW2 (right) mRNAs according to TCGA GBM database. Log rank (Mantel-Cox) test.(F) Bubble chart showing the top 10 hallmark genes enriched in shCtrl GSCs. Size, gene number; color, −log2 (p value).(G) GSEA plot for the PI3K/AKT/mTOR signaling signature from the Hallmark category comparing the shCtrl and shCD97 groups.See also Figure S11.