Let-7c-3p Regulates Autophagy under Oxidative Stress by Targeting ATG3 in Lens Epithelial Cells

Figure 5

Let-7c-3p regulates autophagy by targeting ATG3 in SRA01/04 cells under oxidative stress. (a) Putative let-7c-3p binding site in the 3UTR region of ATG3. (b, c) The expression levels of ATG3 mRNA and protein regulated by upregulated or downregulated let-7c-3p. Real-time PCR and western blot were used to detect the level of ATG3 mRNA and protein expression. ATG3 expression was downregulated when SRA01/04 cells were transfected with let-7c-3p mimics and upregulated with let-7c-3p inhibitors. (d) Relative luciferase level among wild-type ATG3-3UTR, mutated ATG3-3UTR, mimics-NC, let-7c-3p mimics, inhibitor-NC, and let-7c-3p inhibitor. LECs were transfected with let-7c-3p mimics, mimics-NC, inhibitor-NC, let-7c-3p inhibitor, wild-type 3UTR, and mutated 3UTR of ATG3. Luciferase activity was analyzed after 48 h. (e) Western blot was used to analyze the expression level of LC3B II and LC3 I proteins in SRA01/04 cells infected by mimic controls, let-7c-3p mimics, and let-7c-3p mimics with pcDNA3.1-ATG3 under oxidative stress. (f) The schematic of let-7c-3p-regulating autophagy by targeting ATG3 in SRA01/04 cells under oxidative stress ().

(a)

(b)

(c)

(d)

(e)

(f)