Expression of M1/M2 markers in RAW264.7 cells assayed by flow cytometry. The RAW264.7 macrophages with F4/80⁺CD86⁺ phenotype was identified as M1 macrophages, and those with a F4/80⁺CD206⁺ phenotype was designated as M2 macrophages. A: Gating strategy of macrophages. The cell population was analyzed in the P1 gate, and single cell in the P2 was circled using forward scatter-area (FSC-A) and forward scatter-height (FSC-H). Macrophages were identified using F4/80 antibody in the P3 gate. B: Gating strategy of M1 macrophages. Black box displayed the F4/80⁺CD86⁺ M1 cells. C: The percentage of CD86 positive cells was shown in the bar graph. D: Gating strategy of macrophages was the same as (A). E: Black box displayed F4/80⁺CD206⁺ M2 macrophages. F: The percentage of CD206 positive cells is shown in the bar graph. *p < 0.01 compared to the DMEM group.