Supplementary Figure 2: Workflow and Quality Control for sorted single CDP populations.

a Experimental approach for analysis of gene expression in single CD115^- CDPs and single CD115⁺ CDPs using the Fluidigm C1 autoprep system. Violin plots for b number of genes detected in each single cell, c nUMI and d and percentage of mitochondrial genes in the sorted single cell populations. e Bar plot of number of expressed genes in individual sorted single cells. f CMap analysis of enrichment of pDC-signature genes versus cDC-signature genes in CD115^- CDPs and CD115⁺ CDPs. a–f Data are representative for one experiment with 96 single BM CD115⁺ CDPs and 58 single CD115^- CDPs. g Quantification of the cDC and pDC progeny of cultured CD115⁺ CDPs on day 7 of culture with rh-Flt3L. h Flow cytometry analysis of the progeny of sorted, cultured Ly6D⁺CD81^-, Ly6D⁺CD81⁺, Ly6D^-CD81⁺, and Ly6D^-CD81^- CD115^- CDPs on day 7 of in vitro stimulation with rh-Flt3L. g,h Data are representative for three experiments with one replicate per culture condition. i Percentage of Ly6D⁺CD81^-, Ly6D⁺CD81⁺, Ly6D^-CD81⁺, and Ly6D^-CD81^- CD115^- CDPs and SiglecH^-/+ pre-pDCs in the BM. j Percentage, absolute numbers and percentage of apoptotic BM LMPP, CMP, GMP, Ly6D^- CLP, Ly6D^-CD115^- CLP, Ly6D^- pre-pDC, CD115⁺ CDP, Ly6D⁺CD115^- CDP, Ly6D⁺ pre-pDC and Ly6D⁺ pre-cDC. k Percentage, absolute numbers and percentage of apoptotic BM pDCs. l Percentage of proliferating cells among analyzed cell populations. m Flow cytometry analysis of percentage of Annexin V vs. DAPI stained cells among the indicated cell populations. n Flow cytometry analysis of percentage of Annexin V vs. DAPI stained BM pDCs. h–n Mean ± s.e.m. Data are representative for three independent experiments with three mice each.