Figure 2.

LC3B^Y113C mutation reduces autophagy levels in cells. (a) Fluorescence micrographs of HeLa cells transfected with plasmids encoding GFP-LC3B (LC3B WT, LC3B^Y113C, or LC3B^G120A), and treated with DMSO or Torin1 for 2 h. (b) Autophagy levels measured in GFP-LC3B variants treated with DMSO or Torin1. The results were normalized to GFP-LC3B WT treated with DMSO. Autophagy levels were measured as puncta area/total cell area. The calculated results are an average of 3 independent biological experiments. Statistical significance was determined by two-way analysis of variance (ANOVA), *P < 0.05; **P < 0.01; ***P < 0.001. (c) Samples from B were western blotted for GFP and VCL, as loading control. (d) Western blots of HEK293A cells transfected with MYC-LC3B variants (LC3B WT and LC3B^Y113C) and treated with Torin1 or DMSO for 4 h, blotted with anti-MYC antibodies.