Fig. 3: Loss of Mcam promotes macrophage recruitment.

a GSEA of enriched immune response signatures using total mammary cells in cKO mice by MMTV-Cre compared with WT mice. NES, normalized enrichment score. b Percentages of various immune cell populations in WT and cKO mice by FACS analysis. n = 8 mice for macrophages, neutrophils, and DCs, 3 mice for B cells and CD8⁺ T cells, and 6 mice for CD4⁺ T cells. c FACS analysis of macrophage population (F4/80⁺CD11b⁺) in WT and cKO mice. d, e IF images of CD206⁺ (d) and Cx3cr1⁺ (e) macrophages in WT and cKO mammary sections. Scale bar, 20 μm. f, g FACS analysis (f) and their percentage quantification (g) of macrophages in mammary tissues of WT and cKO mice. n = 3 mice in each group. h, i Schematic of co-culture assay and representative images (h) (The left panel was created with BioRender.com.), and quantification (i) of migrating macrophages after WT and cKO CM treatment. Isolated macrophages by FACS in upper chamber were co-cultured with CM from WT and cKO primary MECs in lower chamber. CM, conditional medium. n = 6 biological replicates. Scale bar, 200 μm. Data are means ± SEM. Two-sided Student’s t test was used to evaluate statistical significance. Source data are provided as a Source Data file.