Fig. 3

M2 macrophages confer a partial mesenchymal phenotypic shift of MCF-7. a Fluorescent imaging depicts RFP-MCF-7 cells co-cultured with M0, M1 or M2 macrophages in serum-free hepatocyte maintenance medium (HMM) for 5 days. MCF-7 cells mono-cultured in serum-free HMM on day 5 as the negative control, mono-cultured MCF-7 in their normal growth medium (RPMI 1640 with 10 % serum) on day 5 as the positive control. (Bar = 10 μm). Arrows indicate more elongated, spindle-shaped and mesenchymal-like cells. MCF-7 mesenchymal phenotypic shift in a quantified by the ratio of the midpoint diameter divided by cell perimeter. Data are mean ± s.e.m of 3 independent experiments. **, P <0.01. NS indicates not significant. b MCF7 cells co-cultured with M2 macrophages decreased E-cadherin expression. Immunofluorescence analysis of E-cadherin expression in RFP-MCF7 cells by co-culture with M0, M1 or M2 macrophages. E-cadherin (green), RFP (red), DAPI (blue), Merge (yellow) (Bar = 10 μm). Quantification of the percentage of E-cadherin negative cells in RFP-MCF-7 cells co-cultured with M0, M1 or M2 macrophages. **, P <0.01. 100 cells per field in 10 different fields of each group were quantified. Data are represented of at least three independent experiments. c Flow analysis of E-cadherin expression of RFP-MCF-7 population after 5 days of co-culture with M0, M1 or M2 macrophages. The grey background represents unstained control. The black line indicates co-culture with M0 macrophages. The red line indicates co-culture with M1 macrophages. The blue line indicates co-culture with M2 macrophages. Shown is one of two similar experiments