Fig. 5.
A) Zymosan Phagocytosis in RAW264.7 macrophage. 105 Macrophages were cultured in a medium, irisin (400ng/ml), and LPS (1000ng/ml) alone respectively, or pre-treated for 12 h with irisin, and then LPS were added for 12h. Phagocytosis was allowed to proceed for 30 min. For 30 min at 4 °C, cells were fixed with 4 % paraformaldehyde, followed by a PBS wash. Using a Nikon Eclipse Ti2-E microscope, the plate was analyzed using light microscopy at 400x magnification. B) Dead cell clearance by macrophages. For the dead cell clearance assay, macrophages were co-cultured with a cell tracker labeled (red) thymocytes. Over 90 % of cells become phosphatidylserine positive after incubation with 5 μM dexamethasone. C) scoring of thymocytes engulfed by macrophage. D) Apoptotic thymocytes (red) engulfed by macrophages. Data presented as mean ± SE (n=3). *P< 0.05. Control, medium; Irisin, 400 ng/ml irisin; LPS, 1000 ng/ml LPS; Irisin + LPS: after pretreatment with irisin (400 ng/ml) for 12h, LPS (1000 ng/ml) was added for 12h.