Figure 6
From: Acetylated microtubules are required for fusion of autophagosomes with lysosomes
Vinblastine blocks autophagosome-lysosome fusion on acetylated microtubules. (A) Immunoblotting analysis of LAMP2 levels in HeLa cells treated with different concentrations of nocodazole and vinblastine in the absence or presence of bafilomycin A1 overnight. (B) Colocalization analysis of GFP-LC3 punctate foci with lysosomal marker LAMP2 in HeLa cells stably expressing GFP-LC3 treated with nocodazole or vinblastine in the presence of bafilomycin A1 overnight. LAMP2 was visualized with anti-LAMP2 primary antibody and Rhodamine-conjugated secondary antibody. The overlapping analysis was performed with the ImageJ ColocalizeRGB Plugins. (C) The fraction of GFP-LC3 overlapping with LAMP2 staining as calculated from images shown in (B) with the ImageJ JACoP Plugins. The data shown is the average and standard deviation of ten randomly selected images in a field of 512 pixels × 512 pixels (23.5 μm × 23.5 μm). The significance of differences was determined by Student's t-test.