Figure 6

Effect of chronic alcohol treatment on macrophage MFG-E8 gene expression and efferocytosis. Peritoneal macrophages were exposed to alcohol (50 mmol/L) for 7 d as described in Materials and Methods. After treatments, part of the samples in each treatment group was processed for protein isolation followed by determining MFG-E8 protein with Western blot as described in the Figure 1 legend. The remaining samples were cocultured with apoptotic cells in the 1:4 ratio for 90 min, stained and followed by analysis of macrophages with engulfed apoptotic cells using flow cytometry as described in the Figure 2 legend. (A) Chronic alcohol treatment results in increase in MFG-E8 protein levels in macrophages. A representative autoradiograph from two independent experiments is shown. (B) Chronic alcohol treatment enhances macrophage efferocytosis. A representative histogram of FACS analysis from two independent experiments with triplicate samples in each group.