Extended Data Figure 1. Mycl1 expression within the immune system is restricted to dendritic cells.

a, Shown is the percent of BrdU⁺ cells within each indicated lineage from WT mice 4 hours after BrdU administration. Data are from 2 independent experiments (bars, SD; n = 9 mice per group, one-way analysis of variance (ANOVA) Tukey's post hoc test). b, Shown are single-color histograms of DAPI and Ki67 expression for indicated cell types from the spleen of WT mice. Dotted lines represent isotype control staining. Numbers represent percent of cells in the indicated gate. Data are representative of 3 independent experiments. c, Shown are two-color histograms of CD11c and SiglecH expression for all live BM cells from WT mice, or B220 and MHCII expression for CD11c⁺SiglecH⁺ cells, which were evaluated for DAPI expression (bottom panels). d, Relative Mycl1 mRNA expression was determined by quantitative real-time PCR (qPCR) in pDCs isolated from BM and other indicated cell types isolated from the spleen of WT mice. Shown is a graph of Mycl1 values normalized to Hprt values (bar, SD; n=3 biological replicates per cell type). e, CMP and CDP populations described in Methods Summary were analyzed by gene expression microarray. Shown are transcription factor probe sets with normalized linear expression value in the CDP greater than 100. The horizontal axis indicates the ratio of gene expression in the CMP compared to the CDP, and the vertical axis indicates the ratio of the mean expression in CD8α⁺ cDCs, CD11b⁺ cDCs and pDCs compared to a tissue panel excluding DCs (described in Methods). f, Relative Mycl1 mRNA expression was determined by qPCR for indicated mice. Shown is a graph of Mycl1 values normalized to Hprt values (bar, SD; n=3 biological replicates per cell type). g, Splenocytes from WT and Mycl1^gfp/gfp mice on the C57BL/6 genetic background were stained for analysis, and pDCs (CD11c^intSiglecH⁺), CD8α⁺ DCs (CD11c⁺MHCII⁺CD8α⁺CD205⁺SiglecH⁻), and CD8α⁻ DCs (CD11c⁺MHCII⁺CD8α⁻CD205-SiglecH⁻) were identified. Shown are two-color histograms of CD11c and SiglecH expression, MHCII and CD11c expression, and CD8α and CD205 expression for pre-gated cells as indicated above the plots. Numbers represent percent of cells in the indicated gate. Data are representative of at least 5 independent experiments. h, Relative Myc mRNA expression was determined by qPCR for indicated mice. Shown is a graph of Myc values normalized to Hprt values (bar, SD; n=3 biological replicates per cell type, Student's t-test). *, p<0.05; ns, p>0.05; ND, not detected.