Fig. 7. Cell-intrinsic TNF signaling promotes pDC IFN-I production.

a, Experimental protocol followed to evaluate the role of TNF signaling in pDC activation during MCMV infection. b, Percentages of IFN⁺ cells in splenic pDCs isolated from isotype control (IC)-treated animals vs anti-TNFR2-treated mice. The data are shown for 10 individual animals pooled from 2 independent experiments, with overlay of mean±s.e.m. values. c, Generation of mixed bone marrow chimera (MBMC) mice; CTR, control. Recipient C57BL/6 CD45.1 (5.1) mice were lethally irradiated and then reconstituted with equal proportions of bone marrow (BM) cells isolated from WT 5.1 mice and from indicated CD45.2 (5.2) mice, either WT for CTR MBMC mice or TNFR1/2-KO (TNFRDKO) for TEST MBMC animals. Reconstituted MBMC mice were infected by MCMV and their splenic pDCs examined ex vivo for IFN-I production. d, Measuring the cell-intrinsic role of TNF signaling in promoting pDC IFN-I production. Results are expressed as 5.2/5.1 ratio of the percentages of IFN-I-producing pDCs obtained from MCMV-infected CTR WT versus TEST TNFRDKO MBMC mice. The data are shown for 5 individual animals for each type of MBMC mice, pooled from 2 independent experiments, with overlay of mean±s.e.m. values. For b and d, the statistical analysis was performed using one-sided Mann-Whitney U test. e, Dot plots show representative data of intracellular IFN-I staining in 5.2⁺ versus 5.1⁺ pDCs isolated from the indicated MCMV-infected MBMC mice.