Extended Data Fig. 2: Immune regulation of duct growth during puberty.

a, Quantification of MØs and DCs by FACS in six-week-old CD11c-Cre^T/+Irf8^fl/fl and CD11c-Cre^+/+Irf8^fl/fl mice. Fold-change % CD45⁺ cells was calculated relative to control (n=3 mice, control; n=4, knockout). Mean values are displayed. Error bars, s.e.m. Two-way ANOVA with Sidak correction. *P=0.0142, ns: P=0.6517 (MØ3), P=0.9552 (MØ1/2), P=0.8773 (DC2). b, Carmine-stained whole-mounts from six-week-old CD11c-Cre^T/+Irf8^fl/fl and CD11c-Cre^+/+Irf8^fl/fl mice and branch quantification (n=6 mice, control; n=11, knockout). Error bars, s.e.m. Two-tailed Student’s t-test. Scale bars, 2 mm. c, FACS analysis one day after DT treatment of CD11b-DTR mice (n=3 mice, control; n=4 mice, depletion). Eos: eosinophil, Neut: neutrophil, Mono: monocyte. Means are stated. Error bars, s.e.m. Two-way ANOVA with Sidak correction ****P<0.0001. d, Whole-mounts from CD11b-DTR or WT mice treated with DT (5 weeks) and collected at 6 weeks (n=4 mice, control; n=5 mice, depletion). Error bars, s.e.m. Two-tailed Student’s t-test **P=0.0018. Scale bars, 2 mm. e, FACS analysis of CD11c-Cre^T/+RBPJ^fl/fl (RBPJ cKO) and CD11c-Cre^+/+RBPJ^fl/fl (control) mice at 6 weeks, frequency relative to epithelial cells (CD45⁻CD24⁺) (Two-way ANOVA with Sidak correction, ns: all P>0.9999) and MFI relative to control mean. Two-way ANOVA with Sidak correction ****P<0.0001, ns: P=0.5882 (MHCII), n=0.1659 (CD11c), n=0.9271 (Ly6C) (n=5 mice, controls; n=3 mice, cKO). Error bars, s.e.m. f, Whole-mounts from control and RBPJ cKO mice and quantification of ductal area and branch frequency (n=5 mice, control; n=3 mice, cKO). Error bars, s.e.m. Two-sided Student’s t-tests, P=0.6548 (area), P=0.0538 (branches). Scale bars, 2 mm. g, 3D images from control and RBPJ cKO mice immunostained for MHCII (yellow) and K5 (magenta). Opaque signal rendering as in Fig. 3c. Image representative of 5 mice (control) and 3 mice (cKO). Scale bars, 50 µm.

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