Figure 3

Co-localization of LC3B-II and autophagy effectors in phagocytic cups of IgG-opsonized particles. Non-professional phagocytes were fed with IgG-opsonized beads for 30 min, fixed, and co-stained for F-actin with Phalloidin and for the endogenous LC3B or autophagy adaptors. (A–D) are representative images of cells co-stained for F-actin and p62 (A), LC3B (B), NBR1 (C) or NDP52 (D). The first column represents cells stained for F-actin. The second column represents cells stained for the endogenous p62, LC3B-II, NBR1 or NDP52. The third column shows non-internalized beads stained with an anti-human IgG antibody conjugated with FITC. The fourth column represents merged images of F-actin with LC3B-II or autophagy adaptors and the opsonized latex beads (external and internal beads visualized by DIC). The regions outlined by the boxes are phagocytic cups formed upon the recognition of IgG-opsonized particles by the Fc-receptors. Bars, 10 µm.