The AP-1 transcription factors c-Jun and JunB are essential for CD8α conventional dendritic cell identity

摘要

a Representative flow cytometry plots depict the sorting strategy for pDCsFL (CD11c+ B220+), the cDC FL subsets (CD11c+ I-A/I-E+): cDC1FL (CD24+ cDCFL), cDC2FL (CD172a+ cDCFL), pre-cDCsFL (B220- CD117low-int CD115+ CD11c+ I-A/I-E-), and progenitors (B220- CD117low-int CD115+ CD11c- I-A/I-E-) generated from wild-type bone marrow supplemented with FLT3L for 8d. b RT-qPCR analysis of c-Jun and JunB mRNA expression in progenitors, pre-cDCFL, cDC1FL, cDC2FL, pDCFL. Subsets were obtained according to (a). RNA fold change is shown relative to the progenitor population. c Experimental design. Deletion of c-Jun and JunB in the Mx-Cre mouse model was induced by injection of poly I: C (i.p., 200 µg). Mice were analyzed 6 days after the first injection. d Representative flow cytometry plots show single, live splenic cDCs (CD11c+I-A/I-E+) and CD8α cDC1 (CD8α+ CD11b- cDC) and CD11b cDC2 (CD8α- CD11b+ cDC) subsets from indicated mice. e Frequency and number of splenic CD8α cDC1 and CD11b cDC2 as defined in (d) are shown. f Representative flow cytometry plots depict splenic cDC1 defined as DEC-205+CD11c+ cells in control c-Jun/JunBfl/fl and in c-Jun/JunBΔ/ΔMx-Cre mice. g Skin-draining lymph node (sd-LN) cDCs consisting of migratory DCs (mDCs; CD11c+I-A/I-Ehigh) and resident DCs (rDCs; CD11c+I-A/I-Eint) were analyzed by flow cytometry for the CD8α rDC and CD103 mDC subset in c-Jun/JunBfl/fl and c-Jun/JunBΔ/ΔMx-Cre mice. h Flow cytometric histograms show CD8α or CD103 expression on sd-LN cDCs (CD11c+I-A/I-Eint-high) in the indicated mice. i rDCs and mDCs from (g) are shown as percentage of live, single cells. Data are representative of 2–5 independent experiments. Dots indicate number of individual mice per experimental group. Error bars represent mean ± SEM. Statistical significance was determined by Brown–Forsythe and Welch ANOVA test (b) and unpaired two-tailed Student’s t test (e, i). ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05 and ns > 0.05.