Fig 2: Colony formation and CCK-8 assays showed that H3.3K27M had no obvious effect on proliferation of glioma cells in vitro, and Western blotting showed that H3.3K27M did not change the levels of kinases involved classical proliferation pathways. (A) The overexpression efficiency in U87 and LN229 cells was verified using Western blotting. H3K27M was accompanied by a loss of H3K27me3, and no difference in the total amount of H3 was observed among the three groups. Representative images (B) and statistical analysis (C) of colony formation showed no discernible difference among the vector group, Myc-H3F3A group and Myc-H3.3K27M group in U87 and LN229 cells. (D) The CCK-8 assay revealed no significant effect on the proliferation of the three groups of U87 and LN229 cells. (E) Representative blots of AKT, P-AKT, MEK1/2, P-MEK1/2, STAT3 and P-STAT3 levels in the three groups of U87 and LN229 cells. ns indicates no statistical significance.

Fig 3: IL13Ra2 expression in different BSG subtypes of the TMA cohort. (A) IL13Ra2 expression of BSG tissues from patients of different genders; (B) IL13Ra2 expression of BSG tissues from patients of different ages; (C) expression of IL13Ra2 in different locations of BSG tumor tissues; (D) expression of IL13Ra2 in DIPG and non-DIPG tumor tissues; (E) expression of IL13Ra2 in H3F3A-wt and H3F3A-mt tumor tissues; (F) expression of IL13Ra2 in BSG tissues of different WHO grades (ns p > 0.05, * p < 0.05, ** p < 0.01, **** p < 0.0001).

Fig 4: IL13Ra2 staining extent in BSG tissues. (A) Typical images of multiple fluorescence staining and spectral resolution of BSG FFPE tumor tissues (20×, all scale bars: 50μm); (B) expression of IL13Ra2, H3.3-K27M, Ki67, CD133, CD4, and HLA-1; (C) expression of H3.3K27M in H3F3A-mutant and wild-type tissues.

Fig 5: IL13Ra2 expression in BSGs of the BA cohort. Comparison of IL13Ra2 transcription level in tumor tissues with different H3F3A statuses (A), WHO grades (B), and methylation subtypes (C) in the BA cohort. The linear regression analysis results for IL13Ra2 transcription level and H3.3 (D), Ki67 (E), and CD133 (F) transcription levels in the BA cohort. (G) Kaplan–Meier survival analysis results of the IL13Ra2-high group and IL13Ra2-low group in the BA cohort (* p < 0.05, *** p < 0.001, **** p < 0.0001).