Figure 8

Immunofluorescent and Prussian blue imaging of human macrophage iron deposit polarization and CSF1R status. Representative registered fields of human carcinoma in situ (CIS) and invasive carcinoma where iron deposits are found (INV) stained for iron with Prussian blue (a,b), allograft inflammatory factor-1 (AIF1, c,d), mannose receptor (CD206, e,f), colony stimulating factor 1 receptor (CSF1R, g,h) and the combined immunofluorescent channels (i,j). Each field is 200 µm × 200 µm. Scale bar 50 µm. (k) Number (#) of AIF1⁺, CSF1R⁺, CD206⁺ and iron⁺ macrophages detected per CIS and INV field. (mean+s.e.m. n=18 fields for CIS, n=9 fields for INV, *p < 0.05, ***p < 0.001, ****p < 0.0001, Mann-Whitney test). (l) Fraction of total iron⁺AIF1⁺, iron⁺CD206⁺, and iron⁺AIF1⁺CD206⁺ macrophages detected per CIS and INV field. (m) Fraction of total iron⁺CSF1R⁺AIF1⁺, iron⁺CSF1R⁺CD206⁺, and iron⁺CSF1R⁺AIF1⁺CD206⁺ macrophages detected per CIS and INV field (mean+s.e.m. n=18 fields for CIS, n=10 fields for INV, **p < 0.01, ***p < 0.001, ****p < 0.0001, Mann-Whitney test).