Fig. 1: Relationship of Siglec-H⁺CCR9^loB220^hi cells and pDCs.

a Gating strategy for Siglec-H^– pre-cDC, Siglec-H⁺CCR9^loB220^lo (lo-lo), CCR9^loB220^hi (lo-hi) and CCR9^hiB220^hi pDCs. The indicated populations were sorted from the BM cells of 3 individual C57BL/6 mice and processed for RNA-sequencing. b Principal component analysis performed on normalized counts using all genes. c Expression heatmap of selected TFs, z-score hierarchical clustering by Euclidean distances (normalized counts). d Gene signatures of the sorted populations compared to previously published gene signatures of pre-cDC, cDC, CDP, pDC and pre-pDC³⁹. Distance from the middle of the radar plot indicates the percentage of overlap between genes of sorted populations that showed higher expression than the inter-population median and the indicated gene signatures. e Gating strategy for pDC/cDC output after culturing precursor cells. cDCs were identified as Siglec-H^– MHCII^hi cells and pDCs as Siglec-H⁺CCR9^hiB220^hi cells within CD45⁺CD11c⁺ cells. f Siglec-H^– pre-cDC, lo-lo and lo-hi precursors, and pDC were sorted from Lineage-depleted BM cells of WT mice as shown in (a) and cultured for 3 days on EL08-1D2 stromal cells in Flt3L-containing medium. The percentages of cells with a phenotype of pDC (circles) or cDC (triangles) within CD11c⁺ progeny of the indicated input populations is shown as mean ± SEM (n = 7) and was compared using paired, two-sided t-tests with Holm-Šídák correction for multiple testing. Adjusted p-values: <0.05(*), <0.005(**), <0.0.001(***).