Fig. 3. Arzanol interferes with autophagosome elongation.

Shown are representative microscopy images of HeLa cells stably expressing GFP-LC3-RFP-LC3ΔG (A), or wild-type HeLa cells immunofluorescently labeled for endogenous LC3 (B) or ATG16L1 (C). All cells were starved in serum- and amino acid-free medium for 2 h while incubated with 3 µM arzanol or 10 nM bafilomycin A₁. Scale bars in upper panels are 15.5 µm, scale bars in magnifications are 3.875 µm. (i) Data show the average number of dots per cell from biological triplicates for GFP-LC3, and n = 6 for LC3 and ATG16L1 as mean ± SEM. Digits in bars show the total number of cells quantified using ImageJ software. (ii) Data show the average diameter of dots in nm as mean ± SEM. Statistical analysis was performed using ordinary one-way ANOVA with Tukey’s multiple comparison test, *P < 0.1.