Fig. 5 | Nature Communications

Fig. 5

From: Macrophages orchestrate breast cancer early dissemination and metastasis

Fig. 5

Phenotypic profiling of immune cells in early mammary cancer lesions. a Whole mammary glands from FVB wild-type mice or 14-week-old and 22-week-old pre-malignant MMTV-HER2 mice were analyzed by mass cytometry. viSNE plots were generated from myelomonocytic cells (gating strategy see Supplementary Fig. 4A, B). Results from one representative animal is shown; number of animals per group N = 5; individual experiments N = 2. The three sub-populations were identified as Ly6C+ monocytes and CD206hi and CD206lo macrophages based on their expression levels of Ly6C (b) and CD206 (c). These three populations were then analyzed for their frequency amongst all myelomonocytic cells. df Dot plots show mean±SEM of five animals per group. Heat plots for three individual animals per group with expression levels of selected markers Ly6C (b), CD206 (c), Tie2 (i), and IdU incorporation as a proliferation marker (j) were generated for CD206lo and CD206hi macrophages as identified in the viSNE plots. g, h viSNE plot and quantification of myelomonocytic population in overt MMTV-HER2 tumors (five mice per group). Mammary glands from FVB wild-type mice (k, 22wks), MMTV-HER2 mice at 24wks (l) and overt MMTV-HER2 tumors (m, 26–30wks) were stained against CD206 and F4/80 and CD206 signal intensity in F4/80+ macrophages was quantified; zoom factor in k, l, and m, 2x. Plot n depicts mean±SEM, each dot represents one macrophage; three animals combined. All bars: 10 μm. All statistical testing was done with 95% confidence interval by Mann–Whitney test