Supplementary Figure 5: Migratory DC subsets in MLNs from mock- and MuHV-4 infected mice after HDM sensitization.

(a) Gating strategy for CD103⁺ cDC1, CD11b⁺ cDC2, lung-derived CCR7⁺ DC populations in the MLN of mock or MuHV-4 infected BALB/c mice, HDM sensitized (100 μg) 30 days p.i. and euthanized 2 days later. MLN cells from MuHV-4 infected mice were labelled with CFSE and then mixed with MLN cells from mock-infected mice prior to antibody staining and flow cytometry analysis of a single mix, allowing unbiased comparisons. Debris and doublets were excluded based on FSC and SSC. MOs were excluded based on Ly6c and CD64 expression. Migratory lung-derived CD11b⁺ cDC2 were identified as liveCD11c⁺MHC-II^hiCCR7⁺CD11b⁺ cells. Migratory lung-derived CD103⁺ cDC1s were identified as liveCD11c⁺MHC-II^hiCCR7⁺CD103⁺CD11b^lo cells. Representative flow cytometry plots are shown with the mean frequency of the different cells subsets. (b) Mean fluorescence intensities of maturation markers (CD40, CD80 and CD86, with independent staining and analysis for each of these markers) and MHCII by migratory DCs subsets were compared between the CFSE⁺ (originating from MuHV-4 infected mice) and CFSE^- (originating from mock-infected mice) populations. A reciprocal experiment comparing CFSE⁺ DCs from mock-infected mice to CFSE^- DCs from MuHV-4 infected has been performed as control and gave similar results (not shown).