Fig. 5: TLR7-c-Jun signaling in DCs promotes IL-12 expression.
a–c, UMAP plot of immune cells (CD45+) in B16-F10 tumors shows the distribution of IL-12B in the myeloid cell compartment. Treatment is indicated. Identified cell clusters are color-coded (top right). Fraction of cells positive for IL-12B (b). Cells are assigned to cell clusters (color) identified in UMAP plot. Percentage of type I or II DCs and pDCs positive for IL-12B (c). Classical gating (n = 6 mice per group; data are pooled from two independent experiments). d, Il-12b mRNA expression was quantified in murine BM-DCs generated from wild-type (control) and Tlr7−/− BM. BM-DCs were either pre-treated with IFNα (500 U ml−1) for 6 h and/or stimulated with IMQ (2.5 µg ml−1) over night. (n = 3 mice per group from one experiment). e, Il-12b mRNA expression was quantified by RT–qPCR in BM-DCs generated from c-Junfl/fl and c-Jun∆/∆Mx1-Cre BM. Treatments are as described in d. (c-Junfl/fl: control n = 9, IMQ n = 9, IFNα n = 9, IFNα + IMQ n = 8, c-Jun∆/∆Mx1-Cre n = 6 mice per group; data are pooled from three independent experiments). f, IL-12 protein levels were determined by ELISA in the supernatants of BM-DCs from mice of indicated genotype and treated as described in d. c-Junfl/fl n = 8 mice per group, c-Jun∆/∆Mx1-Cre n = 6 mice per group; data are pooled from two independent experiments. g, Tlr7, c-Jun, and Il-12b mRNA expression was analyzed by RT–qPCR in sorted DCs (CD45+CD64-CD11c+MHC-II+) isolated from B16-F10 tumors implanted in c-Junfl/fl and c-Jun∆/∆CD11c-Cre mice. Tumors were topically treated with IMQ and the mice received IFNα. Tlr7 mRNA: c-Junfl/fl n = 4 or IFNα + IMQ topical n = 6 mice and c-JunΔ/ΔCD11c-Cre n = 6 mice per group, c-Jun mRNA: n = 6 mice per group or c-Junfl/fl control n = 5 mice, Il-12b mRNA: c-Junfl/fl n = 6 mice and c-JunΔ/ΔCD11c-Cre n = 5 mice per group; data are pooled from two independent experiments. Data are plotted as mean ± s.e.m. Dots in c–g represent biological replicates. P values were calculated using one-way ANOVA with Dunnett’s post-test (c) and two-way ANOVA with Sidak’s post-test (d–g).
