Fig. 6: The IMQ antitumor effect depends on c-Jun signaling in DCs.

a–c, Tumor growth was monitored in c-Jun^fl/fl and c-Jun^Δ/ΔMx1-Cre mice (c-Jun^fl/f: n = 9, IMQ topical and oral n = 9, c-Jun^Δ/ΔMx1-Cre: n = 4, IMQ topical and oral n = 8) (a), in c-Jun^fl/fl and c-Jun^Δ/ΔCD11c-Cre (c-Jun^fl/f: n = 19, IMQ topical and oral n = 18, c-Jun^Δ/ΔCD11c-Cre: n = 19, IMQ topical and oral n = 17) (b) and wild-type and Ccl2^−/− mice (C57BL/6J: n = 18, IMQ topical and oral n = 20, Ccl2^−/−: n = 13, IMQ topical and oral n = 19) (c). n in a–c is the number of mice pooled from two (a), four (b) and five (c) independent experiments. d, UMAP analysis was performed on tumor-infiltrating immune cells obtained from mice of indicated genotype, 1 day after IMQ therapy ended. Clustering with CD4, CD8, CD11b, CD64, Ly6-C, Ly6-G and TCRβ. The legend plot shows the identified populations. The fold change of each cluster to the control (c-Jun^fl/fl) is depicted. n = 3 mice per group from one experiment. e–g, Frequency of monocytes, neutrophils and macrophages (e), CD4⁺ and CD8⁺ T cells (f) and pDCs (g) among tumor-infiltrating immune cells in c-Jun^fl/fl, c-Jun^Δ/ΔCD11c-Cre and Ccl2^−/− mice 1 d after IMQ therapy ended. c-Jun^fl/fl: n = 9 (monocytes and macrophages), n = 8 (neutrophils) (e), n = 5 (f), n = 9 (g), c-Jun^Δ/ΔCD11c-Cre: n = 8 (e–g), Ccl2^−/−: n = 9 (monocytes and macrophages), n = 8 (neutrophils) (e), n = 9 (CD4⁺ T cells), n = 8 (CD8α T cells) (f), n = 8 (g); n in e–g is the number of mice pooled from two (e,f) or three (g) independent experiments. h, Protein levels of VEGF-A were measured by ELISA in B16-F10 cells treated with IL-12B (10 ng ml⁻¹) or IMQ (2.5 µg ml⁻¹) for 24 h. IL-12B: n = 6 and IMQ: n = 12 technical replicates of B16-F10 cells per group pooled from two independent experiments. i, B16-F10 proliferation after treatment with IL-12B as described in h. n = 3 technical replicates of B16-F10 cells per group from one experiment. j, Tumor growth kinetics in wild-type mice implanted with B16-F10 melanoma cells. The mice received combination therapy and/or anti-IL-12 antibody (500 µg day⁻¹). n = 4 mice per group or Ig control + IMQ topical and oral n = 6 mice; data are pooled from two independent experiments. k, Quantification of the necrotic area in tumors treated as described in j. Ig control n = 4, anti-IL-12 n = 5; n is the number of tumors pooled from two independent experiments. Data are shown as mean ± s.e.m. Dots represent biological replicates (e–g) and technical replicates (h,i). P values were calculated using unpaired, two-tailed t-tests (h,i,k), one-way ANOVA (e–g) or two-way ANOVA both with Tukey’s post-test (a–c,j).

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