SMAD specific E3 ubiquitin protein ligase 1 accelerates diabetic macular edema progression by WNT inhibitory factor 1

Figure 4 SMAD-specific E3 ubiquitin protein ligase 1 regulated high concentration of glucose/desferrioxamine mesylate-induced tight junction inhibition and autophagy in retinal pigment epithelium cells via WNT inhibitory factor 1. A: ARPE-19 cells were transfected with the SMAD-specific E3 ubiquitin protein ligase (SMURF) 1 or WNT inhibitory factor (WIF) 1 knockdown construct or vector, followed by treatment of the cells with high concentration of glucose or desferrioxamine mesylate, and then the protein level of WIF1 was detected by western blot; B: ARPE-19 cells were transfected with the SMURF1 or WIF1 knockdown construct or vector, followed by the measurement of cellular trans-epithelial electrical resistance; C: ARPE-19 cells were transfected with the SMURF1 or WIF1 knockdown construct or vector, followed by the detection of cell permeability; D: SMURF1, WIF1 or vector knockdown ARPE-19 cells were collected, and the expression of occluding, ZO-1 and claudin-1 was detected by western blot; E: SMURF1, WIF1 or vector knockdown ARPE-19 cells were collected, and the expression of Beclin-1, LC3I, LC3II and p62 was detected by western blot; F: ARPE-19 cells were transfected with the SMURF1 or WIF1 knockdown construct or vector, followed by the immunofluorescence analysis of LC3. The experiments were repeated three times. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. SMURF1: SMAD-specific E3 ubiquitin protein ligase 1; WIF1: WNT inhibitory factor 1; HG: High concentration of glucose; DFX: Desferrioxamine mesylate.