Supplementary Figure 7: Down-regulation of cGAS leads to establishment of immunosuppressive phenotypes.

(a) Tumor growth analysis showed increased proliferation rate of cGAS deficient tumors compared to control tumors. E0771 cells were used for generating tumors with cGAS deficiency. (b–d) FACS analysis for CD11b⁺ Ly6G⁻ Ly6C^high monocytic MDSC, CD11b⁺ F4/80⁺ TAM, and CD11b⁺ Ly6G⁺ Ly6C^low granulocytic MDSC in cGAS down-regulated tumors vs. control tumors. (e,f) The immune cell profile for tumor infiltrating CD4⁺ T cells or CD8⁺ T cells in cGAS deficient tumors vs. control tumors. (g) Knockdown of cGAS decreased the percentage of effector CD8⁺ cytotoxic T cells. Changes in effector CD8⁺ cytotoxic T cells normalized to total CD8 + T cell proportions. (h) Increased level of Treg cells in cGAS deficient tumors, as compared to control tumors. (i) Gene expression analysis of cGAS deficient tumors showed significantly lower level of anti-tumor immunity factors than control tumors. (j) Increased level of tumor promoting factors in cGAS down-regulated tumors vs. control tumors. (k) cGAS expression in tumor biopsies derived from cGAS deficient cells or control cells. The cells expressing shRNA targeting EGFP were used as control in this figure. For panels b–d, n = 4 tumors from 4 different animals. For panels e–h, control: n = 7 tumors from 7 different animals, cGAS-si: n = 10 tumors from 10 different animals. For panels i–k, n = 8 tumors from 8 different animals. Data shown in panel a are means ± SD of n = 8 tumors from 8 different animals mice for each group. For panel g, data is presented as mean ± SD of indicated sample numbers. For panels b–f and h–k, data represent the mean of the above-indicated sample numbers. Samples were compared using two-tailed Student’s t test. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001. ns, not significant.