Fig. 6: Both D2HG and ectopic IDH1 mutant improve therapeutic efficacy of VSVΔ51 in vivo.

a Timeline of the experimental setup for b. b GL261 xenografts were treated with vehicle, VSVΔ51 (i.v., intravenous), D2HG (i.p., intraperitoneal) or a combination. n = 5 mice per group. c Tumor tissues from b were evaluated through immunohistochemistry staining for cleaved-caspase-3 (a marker of cell apoptosis), Ki-67 (a marker of proliferation), VSV-G, and IRF3. Representative images of n = 3. d H&E staining of vital tissues from b. Representative images of n = 3. e Timeline of the experimental setup for f. f Kaplan–Meier survival curve of the mice bearing intracranial GL261-TRE-IDH1(R132H) tumors were treated with vehicle, VSVΔ51 (i.v., intravenous), DOX (i.g., intragastric) or a combination. n = 18 mice per group. NS not significant. g H&E staining of mouse brain from f at day 24 after inoculation. h Immunohistochemistry analysis of cleaved-caspase-3, Ki-67, VSV-G and NKp44 (a marker of NK cells) in tumor tissues from f. Representative images of n = 3. i Percentages of DCs (CD86⁺ CD11⁺) among CD45⁺ cells, CD8⁺ T cells among CD3⁺ cells, Tregs (Foxp3⁺) among CD4⁺ T cells, and central memory T cells (T_CM, CD44⁺ CD62L⁺) among CD8⁺ T cells were analyzed in brain tumor tissues. The function of CD8⁺ T cells was evaluated by measuring granzyme B (GranzB). CD8⁺ T cells with TCR reactivity to the VSV N_52–59 H-2K^b immunodominant epitope were enumerated. n = 5 mice per group. NS not significant. j Splenic lymphocytes extracted from the mice in f were co-cultured with GL261 cells for 48 h. Cell viability of GL261 cells and cytokine secretion of the splenic lymphocytes were detected. n = 3 mice per group. k The survived mice (n = 5) from the combination experiment were rechallenged on day 60. Naive mice of similar age were implanted as controls (n = 5). Statistical significance was determined using two-way ANOVA in b, one-way ANOVA in i, j, or the log-rank (Mantel-Cox) test in f. Data represent the mean ± SD. Scale bar, 50 μm. Source data are provided in the Source Data file.