Extended Data Fig. 3: tDC originate from pro-pDC.

a CD135⁺ BM cells were enriched, sorted as CD45⁺/CD3^-/CD19^-/Ly6G⁻, and prepared for scRNA-seq. Cells with a pDC-specific gene enrichment score of >0.15 were selected and re-clustered using Seurat as shown in Fig. 3d. Violin plots show the expression of the indicated genes in the clusters defined in Fig. 3d. b As in (a) but GSEA of selected pathways from clusters defined in Fig. 3d. Bubble size indicates the normalized enriched score (NES), and color scale depicts False Discovery Rate (FDR). c TdTomato expression in bone marrow progenitor cells from CD300c^TdT mice, gated as in Fig. 3j. One representative experiment. d Imm pDC, pro-pDC and pro-cDC from the BM of CX3CR1^EGFP or hCD2^EYFP CD45.1 mice were purified as described in Fig. 3j and transferred into CD45.2 WT mice. 2- and 4-days post-transfer, recipient mice were analyzed for transferred cells in the spleen. Shown is the number of cells recovered in the spleen of recipient mice (mean + SD). Imm pDC: n = 3 at 2- and 4-days. Pro-pDC: n = 3 at 2-days, and n = 5 at 4-days. Pro-cDC: n = 3 at 2-days, and n = 5 at 4-days. N represents number of mice and independent experiments. e BM cells from CX3CR1^EGFP or hCD2^EYFP CD45.1 mice were CD135-enriched and purified by cell sorting using the gating strategy shown in Fig. 3j. Purified progenitors were adoptively transferred into congenic non-irradiated CD45.2 WT mice and analyzed 4-days later in the spleen of recipient mice. Left panels show post-sort purity of progenitors. Right panels show transferred cells recovered in the spleen of recipient mice, analyzed using the gating strategy described in Fig. 1f. In both cases, lineage includes CD3⁺/CD19⁺/CD335⁺/Ly6G⁺ cells. One representative of ≥ 3 experiments.

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