(A) Experimental scheme for bulk RNA-Seq of BM pDCs sorted from naive mice (red symbols) and BM (green symbols) and lung (blue symbols) pDCs sorted from A. fumigatus–infected mice. i.n., intranasal. (B) Principal component analysis of gene expression in sorted BM pDCs from naive (triangles) and infected mice (circles) and of sorted lung pDCs from infected mice (squares). Each symbol represents a biological replicate. pDCs from 10 mice were pooled for each replicate, and 2 replicates were included in each of 2 experiments, denoted as Expt1 and Expt 2. (C) Number of DEGs for 3 comparisons of 2 pDC subsets, with blue indicating common DEGs and red and green indicating DEGs limited to a single comparison. (D) Differentially enriched KEGG pathways (q < 0.05) in pDCs isolated from infected lungs versus infected BM. Black bars indicate pathways enriched in lung pDCs from infected mice; white bars indicate pathways enriched in BM pDCs from infected mice. (E) Heatmap for 35 selected genes with >2 -fold difference in expression and FDR P < 0.05. Each lane represents 1 replicate from 2 experiments. (F–I) Representative flow cytometry plots (F and G) and quantified MFI (H and I) of IFN YFP and ICAM1 expression in pDCs isolated from naive BM, the BM of A. fumigatus–infected mice, or BM from the lungs of A. fumigatus–infected mice. (A–I) Infection dose: 3 × 10⁷ CEA10 conidia via intratracheal route; analysis 72 hpi. B–E: Data were pooled from 2 independent experiments. H and I: Statistical analysis: Kruskal-Wallis test. See also Supplemental Figure 1.