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. 2021 Apr 12;23(6):444. doi: 10.3892/mmr.2021.12083

Figure 5.

Figure 5.

ATG7 is a direct downstream target gene of miR-654-5p. (A) mRNA expression levels of autophagy-related proteins in NP cells transfected with miR-654-5p mimics. *P<0.05 vs. NC mimics. (B) RT-qPCR was carried out to detect the expression levels of ATG7, ATG16L1 and ULK1. *P<0.05 vs. Normal. (C) A luciferase reporter assay was performed in NP cells cotransfected with miR-654-5p mimics and pmirGLO-ATG7-Wt vectors or pmirGLO-ATG7-Mut vectors. *P<0.05 vs. NC mimics. (D) RNA immunoprecipitation assays were used to further confirm the interaction between miR-654-5p and ATG7. *P<0.05 vs. IgG. (E) mRNA and protein expression levels of ATG7 were detected by RT-qPCR and western blotting in miR-654-5p mimic-transfected NP cells, respectively. *P<0.05 vs. NC mimics. ATG, autophagy-related gene; IDD, intervertebral disc degeneration; miR-654-5p, microRNA-654-5p; Mut, mutant; NC, negative control; NP, nucleus pulposus; RT-qPCR, reverse transcription-quantitative PCR; Wt, wild-type; ULK1, unc-51 like autophagy activating kinase 1.