Figure 2

Acute exposure to alcohol decreases macrophage efferocytosis. Peritoneal macrophages were subjected to treatment with culture medium alone (control) or medium containing alcohol (25 mmol/L) for 18 h as indicated in the figure. After treatments, macrophages were cocultured with apoptotic thymocytes in the 1:4 ratio for 90 min, washed with PBS and processed for staining and analysis of macrophages with engulfed apoptotic thymocytes using fluorescent microscopy and flow cytometry as described in Materials and Methods. (A) Representative photomicrographs of TUNEL-stained samples visualized under a fluorescent microscope. Nuclei of apoptotic cells engulfed by macrophages were demonstrated with bright green fluorescence. Blue fluorescent indicated DAPI-stained macrophage nuclei. Original magnification: 10×. (B) Representative histogram of FACS analysis for quantitatively determining macrophages that engulfed apoptotic cells. (C) Quantitative analysis of efferocytotic ability of macrophages in each treatment group with a flow cytometry—based method. Values are mean ± SEM and represent average of findings from three independent experiments with triplicate samples in each group. **P < 0.01 versus the control group.