Extended Data Fig. 8: Depletion of DMs during involution.
From: Tissue-resident ductal macrophages survey the mammary epithelium and facilitate tissue remodelling
a, FACS of CD11c-DTR mice at 1 day involution, treated with DT or PBS upon forced weaning at 14 d lactation (n=2 mice). b, MØ and DC frequency at 4 d involution by FACS after treatment as in a (n=4 mice, CD11c-DTR; n=10, control). Percent CD45+ cells normalized to controls. Control: CD11c-DTR mice treated with PBS or WT mice treated with DT. DC1, CD11bloCD24hi DCs; DC2, CD11b+CD24int DCs. Error bars, s.e.m. Two-way ANOVA with Sidak correction. ***P=0.0007, ****P<0.0001, ns P>0.9999 (Ly6C+CD11c– MØs), P=0.0941 (DC2). c, Optical sections of CD11c-DTR tissue at 4 d involution after treatment as in (a). Images representative of 3 mice (PBS) and 4 mice (DT). Immunostaining for CC3 (cyan) and MHCII (yellow) and labelling for F-actin (magenta). Scale bars, 100 µm. d, FACS quantification at 4 d involution after treatment with AFS98 anti-Csf1r antibody or isotype control one day prior to weaning, at weaning and at 1 d involution (n=3 mice). Fold-change percent total cells relative to control mean. Monocytes CD64+F4/80+Ly6ChiMHCII–. Error bars, s.e.m. Two-way ANOVA with Sidak correction. *P=0.0212, ***P=0.0008, ns: see Source data. e, H&E staining of tissue from d with alveolar lumen area outlined and quantified (n=3 mice). Error bars, s.e.m. Two-sided Student’s t-test *P=0.0261. Scale bars, 100 µm. f, 3D images of tissue from d immunostained for MHCII (yellow) and CC3 (cyan) and labelled for F-actin (magenta). Images representative of 3 mice. Scale bars, 20 µm. g, Optical sections from a 3D image of Cx3cr1GFP/+ tissue at 6 d involution, immunostained for GFP (yellow) and CC3 (cyan) and labelled for F-actin (magenta). Images representative of 2 mice. Arrows indicate CC3+ DMs. Scale bars, 15 µm.
