Fig 2: AMPK-NF-κB signaling participated in macrophage polarizationBreast cancer cells (MDA-MB231) were treated with metformin 60 μM combined with an AMPK inhibitor (Compound C, CC) or NF-κB inhibitor (BAY-117082, BAY) for 6 h. The supernatant was collected to treat macrophages for 48 h, followed by flow cytometry analysis of CD206, M2 phenotype (A) and CD16, M1 phenotype (B). Data are expressed as mean ± SD, *p < 0.05. DMSO: control; CM: conditioned medium, Met: metformin. Representative flow data shown are from experiments independently performed at least three times.

Fig 3: Metformin treated cancer cells polarized macrophage toward M1 phenotypeTHP-1 cells were stimulated with PMA (200 nM) for 24 h, then incubated with breast cancer (MDA-MB231/MDA-MB453) conditioned medium (CM) with or without metformin (60 μM) for 6 h, followed by flow cytometry analysis to quantify the amount of CD206, an M2 macrophage marker, and CD16, an M1 marker (A, B). Data are expressed as mean ± SD, *p < 0.05. DMSO: control; Met: metformin. Representative flow data shown are from experiments independently performed at least three times.