Fig. 1: Early transcriptional responses of principal accessory cell populations in the lung adenocarcinoma TME to Treg cell depletion.
a, Schematic of the experimental design. b,c, Quantification of Treg (CD4+Foxp3+) one-tailed unpaired t-test P = 12.87, d.f. = 7 ****P < 0.0001 and Tcon (TCRβ+CD4+ and TCRβ+CD8+) cell populations; left, one-tailed t-test P = 0.3799, d.f. = 7, not significant (NS) P = 0.3576; right, one-tailed t-test P= 0.1925, d.f. = 7, NS P = 0.4264, in tumor-bearing lungs 48 h after diphtheria toxin (DT) or PBS (Ctrl) administration. d, Quantification of lung weight in tumor-free and tumor-bearing mice 48 h after DT-induced Treg cell depletion. One-way analysis of variance (ANOVA) followed by Sidak’s multiple-comparisons test. Tumor-free PBS versus tumor-free DT, P = 0.004037, d.f. = 10 NS P > 0.9999; tumor PBS versus tumor DT, P = 0.7450, d.f. = 10, NS P = 0.9787. e, Representative IF staining of Foxp3+ cells in tumor-bearing lungs of Ctrl and DT-treated mice. f, Numbers of upregulated (red) or downregulated (blue) DEGs (P < 0.05) 48 h after DT or PBS administration identified by bulk RNA-seq analysis of the indicated cell subsets. Fib, fibroblasts; Neu, neutrophils; Mac, macrophages; CD4 and CD8, effector CD4+ and CD8+ T cells. g, Representative IF staining of the indicated cell types. h, Quantification of distances between Treg cells and the indicated cell types. One-way ANOVA, alpha = 0.05, followed by Tukey’s multiple-comparison test Treg-Fib tumor-free zone versus tumor nodule, q = 8.041, d.f. = 2544 ****P < 0.0001. Treg-LEC tumor-free zone versus tumor nodule q = 10.08, d.f. = 2544, ****P < 0.0001, Treg-Mac versus tumor-free zone versus tumor nodule q = 17.79, d.f. = 2544, ****P < 0.0001. At least 200 cells were counted in each comparison. Three independent sections per mouse were analyzed. Three and four mice were used in each group in two independent experiments. Data are presented as the mean ± s.e.m. (b–d) (b and c) N = Ctrl-5, DT-4, (d) N = 3 tumor-free PBS, 3 tumor-free DT, 4 tumor PBS, 4 tumor DT. Data are presented as the mean ± s.e.m.
