Fig. 4

Autophagic flux modulates foam cell cholesterol efflux under combined treatment of T0 and MGF. A Peritoneal macrophages incubated with AcLDL were assayed for the puncta of GFP-LC3 (green)/RFP-LC3 (red) during the early stages of autophagy. Scale bar, 20 μm. B Representative digital images of electron microscopy reveal autophagic vacuoles accumulating in the cytoplasm of peritoneal macrophages treated with AcLDL. Scale bar, 50 μm, 1 μm, 500 nm. Quantitation of cholesterol outflow rate with or without chloroquine (CQ) under combined treatment of T0 and MGF in peritoneal macrophages (C) and human THP-1 macrophage treated with AcLDL (D). E Peritoneal macrophages were stained with bodipy (green) and LAMP2 (red) to identify lipophagy in macrophage foam cells under AcLDL-treated conditions. Scale bar, 20 μm, 5 μm. F Expression of ADFP and LAMP2 in AcLDL-treated peritoneal macrophages was determined by western blot with total proteins extracted from cell samples