Figure 2Skin infiltration of pDCs (BST-2+ B220+ CD11cint CD11b-) was analyzed by flow cytometry. Graph shows immune cells as % of live, single cells. (n= 2-15; 1-3 independent experiments). Data information: Data are shown as mean ± SEM. P‐values were calculated by multiple t-test with one-way ANOVA with Bonferroni (A), multiple comparison test. Statistical significance: ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001. See Appendix Table S3 for exact P‐values. Representative flow cytometry plots of skin pDCs 12h after IMQ treatment. Total cell number of skin pDCs defined as in (B) is shown. (n= 5-6: 2 independent experiments) Data information: Data are shown as mean ± SEM. P‐values were calculated by multiple t-test with the Holm-Šídák method (C) multiple comparison test. Statistical significance: ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001. See Appendix Table S3 for exact P‐values. ELISA for CCL2 performed on total back skin treated with IMQ. (n= 5-14; 2 independent experiments). Data information: Data are shown as mean ± SEM. P‐values were calculated by multiple t-test with Tukey (D multiple comparison test. Statistical significance: ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001. See Appendix Table S3 for exact P‐values. BMDCs generated from c-Junfl/fl or c-JunΔ/Δ Mx1-Cre BM were stimulated with IMQ for 12h. CCL2 protein was quantified by ELISA. (n= 7-10; 3 independent experiments). Data information: Data are shown as mean ± SEM. P‐values were calculated by multiple t-test with Kruskal- Wallis (E) multiple comparison test. Statistical significance: ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001. See Appendix Table S3 for exact P‐values. qRT-PCR detection of Ccl2 mRNA in CD11c+MHCII+ cells sorted from IMQ-treated back skin (8h). (n = 5-12; 3 experiments). Data information: Data are shown as mean ± SEM. P‐values were calculated by multiple t-test with Tukey multiple comparison test. Statistical significance: ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001. See Appendix Table S3 for exact P‐values. BMDCs were generated from Tlr7-/-, c-Junfl/fl or c-JunΔ/Δ Mx1-Cre BM and stimulated with IMQ for 4h and/or pre-treated with SP600125 (JNKi, 25µM) for 1h. Ccl2 mRNA was quantified by qRT-PCR. (n = 4-18; 2-4 experiments). Data information: Data are shown as mean ± SEM. P‐values were calculated by multiple t-test with Tukey multiple comparison test. Statistical significance: ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001. See Appendix Table S3 for exact P‐values. H-I. Western blot analysis of Phospho- or total JNK and c-Jun in BMDCs of indicated genotype pre-treated with DMSO (1: 1000) or SP600125 (JNKi, 25 µM) for 1h and stimulated with IMQ (5 µg/mL) for 15 min. Flow cytometry of back skin 12h after intradermal (i.d.) injection of 1 µg rCCL2 and/or IMQ treatment. Analyzed were pDCs (BST-2+B220+CD11cintCD11b-) shown as % of live, single cells. (n = 6; 2 independent experiments). Data information: Data are shown as mean ± SEM. P‐values were calculated by Tukey multiple comparison test. Statistical significance: ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001. See Appendix Table S3 for exact P‐values.