Fig. 4

Claudin5 knockdown promotes autophagy in ESCC cells. A Western blotting was performed to detect LC3 I, LC3 II and Beclin1 protein expression in TE1 cells transfected with Claudin5 siRNA. GAPDH served as the loading control. B transmission electron microscopy (TEM) images of autophagosomes in TE1 cells transfected with Claudin5 siRNA. C mRFP-GFP-LC3-transfected TE1 cells were treated with Claudin5 siRNA, and laser confocal fluorescence microscopy was used to analyze autophagosomes and autolysosomes. Merged image shows yellow puncta for autophagosomes and red puncta for autolysosomes. The puncta were analyzed by ImageJ software. The statistical difference was assessed with one-way ANOVA followed by Dunnett tests in A, B and C. Error bars show the SD from three independent experiments. **p < 0.01, ***p < 0.001, ****p < 0.0001