Figure 7
From: Apoptosis inhibitor of macrophage ameliorates fungus-induced peritoneal injury model in mice
Coating with AIM enhanced debris engulfment by mesothelial cells. (a) Masson’s trichrome stain. Mesothelial cells re-appeared on the surface of the peritoneum on days 21 and 28 in AIM +/+ mice, respectively. The insets are large magnifications of the boxed areas in the respective figures. Scale bars, 100 μm. Findings of electron microscopy of AIM +/+ mice. Sham operation mice: Mesothelial cells were observed on the surface of the peritoneum. Zymosan model AIM +/+ mice on day 14: Macrophages were observed on the surface of the peritoneum. Zymosan model AIM +/+ mice on days 21 and 28: Mesothelial cells were revealed on the surface of the peritoneal membrane (arrow heads). Scale bar, 5 μm. Immunostaining for cytokeratin. Cytokeratin positive mesothelial cells were observed on the surface of the peritoneum of sham operation mice. Cytokeratin positive mesothelial cells re-appeared on the surface of the peritoneal membrane of the AIM +/+ mice (arrow heads) on days 21 and 28. In contrast, mesothelial cells were not observed in the AIM −/− mice. Scale bars, 100 μm. (b) Confocal microscopic images showed phagocytosis by mesothelial cells from both AIM +/+ and AIM −/− mice. Mesothelial cells from AIM +/+ mice had a higher ability of phagocytosis. The arrows indicate phagocytosis. Red: mesothelial cells, Green: dead cell debris, n = 3 dishes for each group, Scale bars, 25 μm. Supplementary Video 3 provides more precise images of phagocytosis by mesothelial cells. (c,d) Cultured mesothelial cells were stained by CellTracker™ Green CMFDA Dye, and mixed with debris labeled by FVD660 with or without rAIM coating. The proportion of engulfment of FVD660 positive dead cell debris by 7-Amino-actinomycin D (7AAD) negative macrophages was assessed. Phagocytosis of mesothelial cells from both AIM +/+ and AIM −/− mice was enhanced by coating the debris with rAIM at 90 min. (C) indicates the flow cytometry using mesothelial cells from AIM −/− mice. n = 3 dishes for each group. Control: Dead cell debris was not incubated, rAIM: Dead cell debris was coated with rAIM.
