Fig. 3: Treatment with iGN increases tumor-infiltrating CD8⁺ T cells and inhibits tumor growth.

a Experimental schedule for evaluating anti-tumor effects of iGN in a therapeutic model. EG7-OVA cells (5 × 10⁵) were subcutaneously (s.c.) transplanted in the right flank on the back of C57BL/6 mice. Five days after transplantation, the mice were injected intradermally with either PBS or iGN (0.4 nmol 1V209/ injection) at the base of the tail thrice every 3 days (at day 5, 8, and 11). On day 14, tumors in each group were subjected to flow cytometry analysis for evaluating tumor-infiltrating CD8⁺ T cells. At day 17, the tumors were subjected to immunohistochemical analysis. b Tumor growth in each group was monitored from day 5 to day 17. Data are presented as means ± SEM; n = 14 mice per group. **p < 0.01, two-way ANOVA test. c Tumors were embedded in paraffin and stained with hematoxylin and eosin (H&E) 17 days after transplantation. Scale bars, 50 μm. d Paraffin sections of tumors were subjected to immunohistochemistry (IHC) analysis using anti-CD8 antibodies 17 days after transplantation. Immunocomplex signals (brown color) developed using 3,3′-Diaminobenzidine (DAB) were visualized; hematoxylin was used as a counterstain. Scale bars, 50 μm. e Percentage of tumor-infiltrating CD8⁺ T cells was quantified 14 days after transplantation. Data are presented as means ± SEM; n = 4 samples per group; each sample represents a pool of tumors obtained from 2–3 mice. *p < 0.05, unpaired t test. f Percentage of IFNγ expression in tumor-infiltrating CD8⁺ T cells was quantified 14 days after transplantation. Data are presented as means ± SEM; n = 4 samples per group; each sample represents a pool of tumors obtained from 2–3 mice. **p < 0.01, unpaired t test. g Percentage of granzyme B (GzmB) expression in tumor-infiltrating CD8⁺ T cells was quantified 14 days after transplantation. Data are presented as means ± SEM; n = 4 samples per group; each sample represents a pool of tumors obtained from 2–3 mice. *p < 0.05, unpaired t test.