rhKAI1 reduces the expression of LPS-stimulated M1 macrophage phenotype-related surface markers in primary mouse bone marrow-derived macrophages (BMDM) and RAW 264.7 mouse macrophage-like cells. Cells were stimulated with various concentrations of rhKAI1 for 24 h or pre-treated with or without rhKAI1 for 2 h, following by 100 ng/ml LPS treatment for 24 h. (A) Representative microscopy images of the cell. Cell morphology was observed using an inverted-phage contrast microscope, and acquired at 100× magnification. (B) Representative flow cytometry plots of F4/80+CD86+ in BMDM cells. (C) Quantification for M1 phenotype are displayed as the percentages of F4/80+CD86+ cells from CD11b-PE-gated cells in BMDM. (D) Quantitative mean fluorescence intensity (MFI) of F4/80+CD86+ cells in BMDM. (E) Representative flow cytometry plots of F4/80+CD86+ and F4/80+CD80+ in RAW 264.7 cells. (F) Quantification for M1 phenotype are displayed as the percentages of F4/80+CD86+ cells and F4/80+CD80+ cells from total live-gated cells in RAW 264.7 cells. (G) Quantitative MFI of F4/80+CD86+ cells and F4/80+CD80+ cells in RAW264.7 cells. Data are expressed as the mean ± SD (n = 5). ***P < 0.001 compared to control. #P < 0.05, ##P < 0.01 and ###P < 0.001 compared to LPS-stimulated cells. (H) Representative images were acquired using a confocal laser scanning microscopy. Confocal images show the expression of CD86 (green) and DAPI (blue). Scale bar = 50 μm.
