Fig. 2: MEOX2 and SRGN are essential for GSCs proliferation, self-renewal, and stemness in CL and MES GSCs respectively.

a, b Cell viability of indicated GSCs with shCONT, shMEOX2, or shSRGN measured by CellTiter-Glo assay (n = 6 per group). Data are presented as mean ± SD. c, e The number of spheres formed by CL and MES GSCs transduced with shCONT or shRNAs targeting MEOX2 or SRGN was quantified (n = 6 per group). Data are presented as mean ± SD. Box plots show median (center line), 25th–75th percentiles (box bounds), and whiskers extend to minimum and maximum values within 1.5 times the interquartile range. ns: non-significant. d, f In vitro limited dilution assay of CL and MES GSCs transduced with shCONT, shMEOX2-1/2 or shSRGN-1/2, respectively (n = 30 per group). χ2 test. g, j Heatmap of SOX2 and ID1 expression in MEOX2 and SRGN knockout RNA-seq data. h, k Quantitative RT-PCR assessment of SOX2 and ID1 mRNA levels in CL GSCs (3028, RKI) and MES GSCs (2907, 839) expressing shCONT, shMEOX2-1/2 or shSRGN-1/2, respectively. Data are presented as mean ± SD. n = 3 per group. i, l IB analysis of SOX2 and ID1 levels in CL GSCs (3028, RKI) and MES GSCs (2907, 839) transduced with shMEOX2, shSRGN, or control shRNA, respectively. m, o Bioluminescence imaging and quantification of orthotopic xenografts derived from luciferase-expressing 3028 (CL) and 2907 (2907) GSCs transduced with shCONT, shMEOX2, or shSRGN. Data are presented as mean ± SD. n = 3 per group. n, p Kaplan–Meier survival curves of mice bearing intracranial tumors derived from 3028 (CL) and 2907 (MES) GSCs (n = 6 per group) from the respective groups above. Statistics: a, b, c, e, m, o one-way ANOVA with Dunnett’s multiple-comparison test. d, f χ² test for two-group comparison. h, k Unpaired Student’s t test for two-group comparison. n, p Log-rank test. Images: i, l Representative blots (n = 3). Source data are provided as a Source Data file.