Figure 3. F4/80⁺Lyve-1⁺ cells are enriched for CD206 expression.

(A) Violin plot of Mrc1 expression in the single-cell RNA-seq dataset shown in Figure 2. (B) Mammary glands from 10-week-old mice (n = 4) were assessed for CD206 expression in CD45⁺CD11b⁺F4/80⁺Lyve-1^- and CD45⁺CD11b⁺F4/80⁺Lyve-1⁺ cells by flow cytometry. (C) Mammary glands were harvested from 6-week-old mice (n = 5, three images/localization), immunostaining was performed for F4/80 and CD206, and the localization of single- and double-positive cells associated with TEBs was examined. Representative images of F4/80⁺CD206^– (i, arrowheads) and F4/80⁺CD206⁺ (ii, arrowheads) cells are shown. (D, E) Representative images of F4/80⁺CD206⁺ cells in the adipose stroma and fibrous capsule of the mammary gland. Representative images show F4/80 (iii) and co-staining (iv). The yellow line in panel (E) shows the margin of the mammary gland. Insets show higher magnification. (F) Flow cytometry analysis showing F4/80^hi and F4/80^int cells in the mammary gland and CD206 expression in each population. (G) Flow cytometry analysis of F4/80 expression on Lyve-1^– and Lyve-1⁺ cells. n = 4 mice analyzed.

Figure 3—figure supplement 1. F4/80⁺Lyve-1⁺ cells are enriched for CD206 expression.

(A) Gating of flow cytometry for Lyve-1 expression in CD45⁺CD11b⁺F4/80⁺CD206^– and CD45⁺CD11b⁺F4/80⁺CD206⁺ cells. (B) Gating of flow cytometry for CD206 expression in CD45⁺CD11b⁺F4/80⁺Lyve-1^– and CD45⁺CD11b⁺F4/80⁺Lyve-1⁺ cells.