Increases of C/EBPβ expression and bindings at Zeb2 −165 kb in CDP by IL-6. (A) RNA-seq was performed on CDPs cultured for 6 h with Flt3L alone or with added IL-6 (25 ng/ml) as described in Fig. 3. Shown is a volcano plot of differentially expressed genes as fold change (FC) induced by IL-6. Selected genes with greater than threefold changes are highlighted in red (increased) or blue (decreased). (B) Shown are genes from A with greater than threefold decrease (blue) or greater than threefold increase in expression (red) in response to IL-6 in CDPs after 6 h. (C) Heatmap of top 101 differentially expressed genes from A clustered on log₂ expression value. (D and E) Characterization of Hoxb8-transduced BM progenitor cell line. (D) FACS analysis of Hoxb8 cells and BM progenitors. (E) Intracellular expression of C/EBPβ in Hoxb8 cells cultured with or without IL-6 (25 ng/ml) for 22 h. Isotype control (IC) is a mouse IgG2a. (F) cDC differentiation from Hoxb8 cell line cultured with or without IL-6 (25 ng/ml) for 7 d. (G and H) CUT&RUN analysis for C/EBPβ bindings to the Zeb2 locus in M-CSFR⁺ Hoxb8 cells cultured in Flt3L with or without IL-6 (25 ng/ml). (G) A snapshot of UCSC genome browser (chr2:44,912,814-45,315,672). Zeb2 −165 kb enhancer is highlighted in yellow. (H) A scatter plot shows total tag counts normalized to IC experimental group in the Zeb2 −165 kb enhancer. Peak calling was performed with HOMER command findPeaks.