Fig. 5: Mcam loss recruits macrophages through Il4-Stat6 axis.

a Heat map showing differentially expressed cytokine genes based on RNA-seq data in cKO mice by MMTV-Cre compared with WT mice. Differentialy expressed genes are labelled in red. b qRT-PCR analysis of expression levels of cytokines in MECs from WT and cKO mice. n = 3 biological replicates in each group. c Western blotting showing Stat6 and p-Stat6 levels in WT and cKO mammary tissues. n = 3 mice in each groups. d, e Representative images (d) and their quantification (e) of migrated primary macrophages incubated with Il4 antibodies or their control (anti-IgG) after treatment with conditional medium (CM) of WT or cKO MECs. n = 6 biological replicates in each group. Scale bar, 100 μm. f, g IF images of CD206⁺ macrophages (f) and their statistical results of CD206⁺ macrophages (g) in WT and Mcam cKO mammary tissues treated with IgG or Il4 antibodies. WT and cKO Mice were treated with IgG (10 μl) or Il4 antibodies (10 μl) orthotopically at 5 weeks of age every other day for a week before harvesting the mammary tissue. Scale bar, 20 μm. n = 3 mice were treated for each groups. 5 fields in each sample for statistics. h, i IF images of Cx3cr1⁺ macrophages (h) and their statistical results (i) in WT and Mcam cKO mammary tissues treated with IgG or Il4 antibodies. Scale bar, 50 μm. 3 mice were treated in each group. 5 fields in each sample for statistics. j, k Representative images (j) and quantification of filled fat pad (k) of whole-mount-stained mammary tissues in WT and cKO mice treated with IgG or Il4 antibodies. n = 6 in WT group and n = 5 in cKO group. Scale bar, 5 mm. WT and cKO Mice in Figure h–k were treated with IgG (10 μl) or Il4 antibodies (10 μl) orthotopically at 5 weeks of age every other day for a week before harvesting the mammary gland. 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.