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. 2014 Nov 14;11(1):88–99. doi: 10.4161/15548627.2014.984277

Figure 3.

Figure 3.

Chloroquine-mediated LC3 recruitment is independent of PtdIns3P and autophagy receptor proteins. (A) Time-lapse microscopy of GFP-LC3 and 2xFYVE-mCherry on entotic corpse vacuoles following treatment with CQ (100 μM). Arrows indicate GFP-LC3 lipidation onto vacuole. Bar = 2 μm. (B) Confocal images of entotic corpse vacuoles treated with CQ (100 μM) with or without LY290004 (25 μM). Arrows indicate GFP-LC3 lipidation onto entotic vacuole, arrowheads indicate 2xFYVE-mCherry-positive vesicles. Bar = 2 μm. (C) Quantification of GFP-LC3 recruitment to LAMP1-positive entotic vacuoles with or without LY290004 (LY 25 μM) or wortmannin (WM, 200 nM); data are mean ± SEM from 3 independent experiments; NS, not significant. (D and E) Entotic corpse vacuoles treated with CQ (100 μM) for 1 h and immunostained for (D) SQSTM1 or (E) CALCOCO2. Arrows indicate autophagosomes with colocalized LC3 and SQSTM1 or CALCOCO2. Bar = 4 μm. See also Figure S4.