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

Figure 4.

Figure 4.

Chloroquine-mediated noncanonical LC3 lipidation is dependent on water flux. (A) Confocal images of LAMP1-GFP fluorescence in MCF10A cells before and after treatment with CQ (100 μM) with or without phloretin (180 μM). Insets show vesicles with LAMP1-GFP. (B) Quantification of LAMP1-GFP-labeled vesicle size; P < 0.01 **** by one-way ANOVA. (C) Confocal images of LysoTracker Red in MCF10A cells before and after treatment with phloretin with or without CQ for 15 min. (D) Confocal images of GFP-LC3 and LAMP1 immunostaining on entotic corpse vacuoles in MCF10A cells following treatment with CQ with or without phloretin (180 mM) or HgCl2 (15 mM). Arrow indicates GFP-LC3 lipidation onto a vacuole. Bar = 5 μm. (E) Quantification of GFP-LC3 lipidation onto LAMP1-positive entotic corpse vacuoles as in (D), data mean ± SEM from 3 independent experiments; P < 0.001 ****. (F) Western blot analysis of LC3 in MCF10A cells treated with phloretin, CQ or both for 1 h. Quantification of LC3-II/LC3-I graphed below. See also Movie S2.