Fig. 3
Cytotoxic activity of mouse macrophages against cancer cells. (A) Mouse peritoneal macrophages were polarized to M1 phenotype by preincubation with LPS (100 ng/mL) + IFN-γ (40 ng/mL) or CA2-2 (0.05 µM) alone. EAC cells were incubated with polarized macrophages for 24 or 48 h. Macrophage density was 3.6 × 104 cells/well; EAC cell density was 3 × 103 cells/well. Cell viability was detected by the MTS method. (B-D) The cytotoxic activity of RAW 264.7 macrophages against different triple-negative breast cancer cells via paracrine manner. Culture medium (CM) was obtained from RAW 264.7 macrophages treated with CA2-2 (0.05 µM) alone or with CA2-2 (0.05 µM) and LPS (10 ng/mL) + IFN-γ (20 ng/mL) for M1 polarization for 24 h. Control macrophages were treated with DMEM medium only for M0 phase. Cancer cells were incubated with CM for 24 and 72 h. Data are expressed as the means ± SEM (n = 5). *p < 0.05, ***p < 0.001 compared to control