Fig. 4. CHEK2 expression in tumor cells is inversely associated with enhanced antigen presentation on tumor cells, in human and mouse gliomas.

a UMAP (left) and violin plot (right) of the gene signature scores of antigen processing and presentation pathway in n = 6,863 tumor cells high and low CHEK2 expression from 28 human glioblastoma tumor samples. b Violin plot of the gene signature scores of T-cell-mediated cytotoxicity pathway in n = 94 T cells from high CHEK2 vs low CHEK2 expressing samples. The scRNA-seq data was used from the study published by Neftel et al.²⁸. For (a and b), the p value represents two-tailed Mann–Whitney test; whiskers represent minimum and maximum values, the white dot inside the box represents the median and the box extends from the 25th to 75th percentiles. c Schematic showing the assay design to test the ability of Chek2 KO cells to promote OT-I CD8+ T-cell activation, assessed by cell proliferation (expansion index). d Flow cytometry analysis showing surface expression of MHCI-SIINFEKL on GL261 Chek2 KO and non-targeting control (NTC) clones, at the basal level and upon stimulation with IFNγ for 48 h. The histogram shows mean ± SD of one representative experiment of 3 independent experiments. p = 0.0059 by two-sided t test. e Four samples-NTC, Chek2 KO, NTC + IFNγ and Chek2 KO + IFNγ were cultured in individual well of the 6-well plates in triplicates. Each of the replicates of these treatment groups were cocultured with CD8+ T cells and OT-I CD8+ T cells independently (N = 3/experimental condition). WT and OT-1 CD8+ T-cell proliferation in all 24 samples was assessed by eFluor 450 fluorescence dilution individually. For NTC + IFNγ and Chek2 KO + IFNγ groups, the respective clones were stimulated with IFNγ for 48 h prior to culturing with WT CD8⁺ T cells or OT-I CD8⁺ T cells. Histograms represent mean ± SD of N = 3 replicates/condition. p values by two-way ANOVA. Source data for d, e are provided as a Source Data file.