Fig. 7

Methyl- β-cyclodextrin efficiently rescues impaired autophagosome-lysosome fusion by U1866A and bafilomycin A1. A RPE1 cells were treated with U18666A (2 μg) or bafilomycin A1 (25 nM) for 12 h in a serum-starved medium. Cells were further incubated in the presence or absence of MβCD (20 μM) for additional 24 h without changing the medium. Cells were then fixed with 4% paraformaldehyde and immunostained for LAMP2 and LC3. Scale bar, 10 μm. B The quantification of GFP-LC3 puncta overlapping with LAMP1 from 30 transfected cells using ImageJ software was determined. Bar graph represents the mean ± SD (n = 3 experiments). #P < 0.05, Student's t-test (test groups were compared with control). *P < 0.05 Student's t-test (respective test groups were compared in the presence or absence of MβCD)