Fig. 1 ∣. Transient NFIL3 expression drives cDC1 specification.

a, Schematic of the Nfil3^GFP reporter mice. Filled and open boxes denote coding and noncoding exons of Nfil3, respectively. b, Representative flow cytometry plots showing GFP–NFIL3 expression in MDPs, CDPs and pre-cDC1 progenitors from the BM of Nfil3^GFP/GFP mice. The percentage of each cell type expressing GFP–NFIL3 is shown. c, Nfil3 transcripts normalized to Gapdh, measured by quantitative PCR with reverse transcription (RT–qPCR), in GFP–NFIL3⁻ CDP, GFP–NFIL3⁺ CDP and pre-cDC1 from Nfil3^GFP/GFP mice. Data are pooled from three independent experiments (n = 3 for each cell type). d, Frequency of splenic cDC1 in Nfil3^fl/fl, Nfil3^fl/fl Itgax-cre and Nfil3^fl/fl Vav-cre mice. Data are pooled from five independent experiments (n = 12 for Nfil3^fl/fl, 9 for Nfil3^fl/fl Itgax-cre and 5 for Nfil3^fl/fl Vav-cre mice). Nfil3^fl/fl versus Nfil3^fl/fl Itgax-cre, P = 0.9067. e, Representative flow cytometry plots showing splenic cDC1s and cDC2s in Nfil3^fl/fl, Nfil3^fl/fl Itgax-cre and Nfil3^fl/fl Vav-cre mice. Percentages of each cell type are indicated. f, CUT&RUN and ChIP–seq tracks display NFIL3 binding around the Zeb2 locus in GFP–NFIL3⁻ BM cells, GFP–NFIL3⁺ BM cells, the NFIL3-expressing Hoxb8 cell line or T cells, visualized with the UCSC genome browser. g, Homer NFIL3 motif. h, FIMO analysis depicting P-values of the three NFIL3 binding sites in the −165 kb Zeb2 enhancer. The Homer NFIL3 motif (shown in g) is used in FIMO analysis. Red residues indicate core NFIL3 motif. Data in c are mean±s.d. Centre values in d indicate median. NS, not significant. Brown–Forsythe and Welch ANOVA with Dunnett’s T3 multiple comparisons test is used in d.