Extended Data Fig. 4: LiMacs proliferate during the lactation period in mice.
From: Lactation-associated macrophages exist in murine mammary tissue and human milk
a, UMAP showing proliferating cells (in red) of scRNA-seq data of myeloid cells (CD11b+ and/or CD11c+) and dot plot showing expression of proliferation-associated genes in lactating mammary glands (at day 7 pp) of wild-type mice. Dot size indicates percentage of cells expressing the gene; color indicates the average gene expression level per cell. Related to Fig. 2. b, Polar volcano plot showing differentially expressed genes between liMacs at days 8, 11 and 14 pp, analyzed by the limma voom method. Coordinates correspond to relative mean expression values per group. 86 genes out of 8052 resulted differentially expressed, of which 46 were specifically upregulated in day 8 pp (adjusted p values < 0.01). 30 genes with top p values are labeled. Related to Fig. 2. c, Representative immunofluorescence image and a bar graph (±SEM) showing percentage of Ki67+ liMacs in lactating mammary glands from Cx3cr1GFP/+ dams (day 8–10 pp). Ki67 (magenta), SMA (yellow), Cx3cr1-GFP (green) and DAPI (blue). Insets are magnifications showing single stainings of the outlined regions. N = 4 mice. Scale bar: 75 µm, inset 18.75 µm. d, Violin plots showing total cell counts of Ly6Chi monocytes, neutrophils and eosinophils in lactating mammary glands (day 8–14 pp) of Csf1rfl/fl and Cd11cCreCsf1rfl/fl dams. Pooled data from 5-6 mice analyzed in 4 independent experiments. Two-tailed Mann-Whitney test was performed, ns = not significant. Related to Fig. 4. e, Violin plots showing total cell counts of Ly6Chi monocytes, neutrophils, eosinophils, B cells and DCs in lactating mammary glands (day 7–14 pp) of control (isotype control antibody-treated or untreated) or CSF-1 antibody (Ab)-treated (2-3 times) from wild-type dams. Pooled data from 11 mice analyzed in 5 independent experiments. Two-tailed Mann-Whitney test was performed, ns = not significant. Related to Fig. 4.
