Extended Data Fig. 3: Characterization of DC progenitors and splenic DC populations.

(a) Expression levels of selected TF-encoding genes in different stages of DC development assessed by microarray. (b) IRF8 Expression across different stages of DC development in WT mice as determined by intracellular staining. Numbers are geometric MFI. Data shown are representative of two similar experiments. MDP, monocyte-dendritic cell progenitors. (c) Frequencies of the indicated DC progenitors from WT, H/+, and H/H mice as percentages of lin⁻ SiglecH⁻ BM cells. Data are pooled from three independent experiments (n = 5 for WT, 3 for H/+, and 4 for H/H mice). Statistical significance was determined by one-way ANOVA. (d) Frequencies of splenic pDCs (gated as B220⁻ SiglecH⁺ splenocytes) from WT, H/+, and H/H mice. Data are pooled from three independent experiments (n = 9 for WT, 5 for H/+, and 7 for H/H mice). Statistical significance was determined by one-way ANOVA. (e) ZBTB46 expression in the indicated cell types from H/H spleens measured by intracellular staining. B cells serve as a negative control for ZBTB46 expression. (f) In vitro cross-presentation assay with different DC populations. Shown are representative flow cytometry plots depicting CellTrace Violet-labeled OT-I CD8 T cells co-cultured with the indicated DC subsets and HKLM-OVA antigen for three days (pre-gate: CD45.2⁻ CD45.1⁺ CD8a⁺ CD4⁻ Va2⁺ cells). Divided OT-I cells indicate successful cross-presentation and T cell activation. (g) Frequencies of divided OT-I cells from in vitro cross-presentation assays. Data are pooled from two independent experiments. (h) Overlaid flow cytometry plots showing splenic cDC1s (cyan), cDC2s (red), and hybrid DCs (purple) before (top panel) and after (middle and bottom panels) culturing in the specified conditions. The data presented are representative of three similar experiments. Data in c and d are presented as mean values +/− SD.