IL-6 increases Cebpb expression in CDPs. (A) RNA-seq was performed on CDPs cultured for 6 h with Flt3L alone or with added IL-6. Shown is a volcano plot of differentially expressed transcription factors as the fold change (FC) induced by IL-6. Selected genes with greater than twofold change are highlighted in red (increased) or blue (decreased). (B) Heatmaps of the top 36 differentially expressed transcription factors from A clustered on (upper panel) Z-score or (lower panel) log₂ expression value. (C) Intracellular expression of C/EBPβ and IRF8 in sort-purified CDPs treated in vitro with Flt3L (blue) or Flt3L with 25 ng/ml IL-6 (red) for 20 h. IC indicates staining using isotype control antibody. (D) Intracellular expression of C/EBPβ and IRF8 in CDPs from BM of Zbtb46^egfp/+ mice analyzed 7 d after tumor inoculation i.p. as indicated. (E) Bar-scatter graphs show the geometric mean fluorescence intensity (MFI) ± SD for experiments shown in E for C/EBPβ (red) and IRF8 (blue). (F) Sort-purified CDPs were transduced in vitro with retroviral vectors (RV) encoding LAP, LIP, or IRF8, cultured in Flt3L for 4 d, and analyzed for FACS. Shown are Thy1.1⁺ B220⁻ SiglecH⁻ cells for expression of MHCII, CD11c, CD24, and Sirpα. Results are representative of four independent experiments. (G) Scatter plots with error bars for cells described in F for total MHCII⁺ CD11c⁺ cells (left panel) or for MHCII⁺ CD11c⁺CD24⁺ Sirpα⁻ cells (right panel; average % ± SD). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (Student’s t test).