Extended Data Fig. 7: Analyses of intermediately optimized Irf8 + 32 kb enhancers.

(a) Diagrams and sequences of the different Irf8 + 32 kb enhancer alleles generated in this study. (b) Retroviral reporter activities of various Irf8 + 32 kb enhancer constructs in cDC1s obtained from Kit/Flt3L-cultured BM. Shown are enhancer-driven GFP expression in transduced cDC1s (pre-gate: B220⁻ CD11c⁺ MHCII⁺ CD24⁺ Sirpa⁻ Thy1.1⁺ cells). Numbers are GFP geometric MFI. Data shown are pooled from two independent experiments (n = 3 for WT mice). Statistical significance was determined by one-way ANOVA with Dunnett’s multiple comparison test. (c) Representative flow cytometry plots of splenocytes stained for cDCs (pre-gate: B220⁻ SiglecH⁻ CD11c⁺ MHCII⁺ splenocytes). (d) Frequencies of the indicated DC subsets as percentages of B220⁻ SiglecH⁻ CD11c⁺ MHCII⁺ cDCs. Data are pooled from six independent experiments. Statistical significance was evaluated using one-way ANOVA with Dunnett’s multiple comparisons test. (e) Representative flow cytometry plots of CD11c-enriched splenocytes stained for cDCs (pre-gate: B220⁻ CD11c⁺ MHCII⁺ splenocytes). (f) Frequencies of the indicated DC subsets as percentages of B220⁻ CD11c⁺ MHCII⁺ cDCs. Data are pooled from two independent experiments (n = 4 for WT and I/I mice). Statistical significance was determined by unpaired, two-tailed Student’s t test. (g) Representative histograms showing intracellular IRF8 levels in DC subsets gated in (e). Numbers are geometric MFI. (h) IRF8 geometric MFI in cDC1s gated in (e). Statistical significance was determined by unpaired, two-tailed Student’s t test. Data shown are pooled from two independent experiments (n = 4 for WT and I/I mice). Data in b, d, f, and h are presented as mean values +/− SD. P values are indicated above the graphs.