Fig. 3

TAMs polarized in OCM/AS enhance the metastatic potential in EOC. (A-D) EOC cells were seeded in the upper chamber, while M0 MΦs, OCM-MΦs, or no cells were seeded in the lower chamber of the transwell plates and incubated for 36 h. The stained cells were counted in four randomly selected fields. Representative images and quantitative results of cell migration and invasion are shown (n = 4). Scale bar: 50 μm. (E-F) The ovarian cancer cell lines OVKATE and OVSAHO cells were incubated with either the control or the conditioned medium (CM) from various macrophage subsets (M0 MФs, OCM-MФs, M1 MФs, or M2 MФs). Subsequently, the viability of OVKATE and OVSAHO cells was assessed by the XTT cell proliferation assay. All the experiments were performed in triplicate (n = 3). (G-H) The spheroid formation capacity of EOC cells alone or co-cultured with control MΦs or TAMs was determined by spheroid formation assays. The size of the spheroids was quantified as shown (n = 9). Scale bar: 50 μm. The data were shown as the mean ± SEM and were analyzed by unpaired Student’s t-tests. The number of independent experiments was n = 9, *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001