Fig. 1.

O-GlcNAc negatively regulates autophagic flux. a – The 5637-GFP-LC3 cells were treated with different doses of DON. GFP-LC3 fluorescence was captured with fluorescence microscopy 16 h later. b – The 5637-GFP-LC3 cells were treated with DON (50 μM). GFP-LC3 fluorescence was captured with fluorescence microscopy at different time points. c and d – The expressions of O-GlcNAc (RL2), LC3 I, LC3 II and p62 in 5637 cells described in (a) and (b) were determined using western blotting assays. e – The 5637-GFP-LC3 cells were treated with different doses of TG. GFP-LC3 fluorescence was captured with fluorescence microscopy 16 h later. f – The 5637-GFP-LC3 cells were treated with TG (10 μM). GFP-LC3 fluorescence was captured with fluorescence microscopy at different time points. g and h – The expressions of O-GlcNAc (RL2), LC3 I, LC3 II and p62 in the cells described in (e) and (f) were determined using western blotting assays. The average number of GFP-LC3 puncta was calculated in 200 cells. Data in the histograms are shown as the means ± SD. Scale bar: 20 μm. *p < 0.05, **p < 0.01, ***p < 0.001