Figure 1.

T-MPs polarize macrophages toward M2 phenotype under normoxia. (A) Real-time PCR analysis of arginase1 mRNA expression in monocytes, bone marrow-derived M0, M2 and M1 macrophages, primary peritoneal macrophages and Raw264.7 cell line treated with H22-MPs for 24 h under normoxic condition. (B) The expression of CD206, IL-10 and CD301 in M0 and M2 macrophages treated with H22-MPs for 24 h under normoxic condition was analyzed by flow cytometry. (C) The expression of NOS2, TNF-α, IL12p35 and IL12p40 in M1 macrophages treated with H22-MPs for 24 h under normoxic condition was detected by real-time PCR. (D) Flow cytometry analysis the expression of TNF-α and IL-12 in M1 macrophages treated with H22-MPs. (E) The expression of iNOS in M1 macrophages treated with H22-MPs for 24 h under normoxic condition was analyzed by protein gel blot. (F) M0 macrophages were treated with T-MPs from H22 tumor cells under UV irradiation or hypoxic condition. IL-4 was used as a positive control. (G) The arginase1 expression in M0 macrophages treated with MPs generated from liver and spleen cells under normoxic and hypoxic condition. MPs generated from liver and spleen cells were added to M0 macrophages for 24 h, and then the expression of arginase1 was analyzed by real-time PCR. (H) MPs or lysate derived from H22, LLC, CT26, B16 and 5 mM lactic acid were added to M0 macrophages for 24 h, the expression of arginase1 was detected by real-time PCR. All experiments were performed at least twice. The histogram bars represent the expression level of three biological replicates, displayed as means±s.e.m. *p <0.05, ***p <0.001. NS, not statistically significant.