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. 2021 Jun 2;48(1):144. doi: 10.3892/ijmm.2021.4977

Figure 3.

Figure 3

miR-30a-5p directly interacts with the 3′UTR of Beclin-1. (A) The potential binding sites of miR-30a-5p and the 3′UTR of Beclin-1. (B) 293T cells were co-transfected with the pGLO-Beclin-1-WT or pGLO-Beclin-1-Mut plasmid and miR-30a-5p mimic or miR-30a-5p mimic-nc, and luciferase activity was detected (n=3). (C) HK-2 cells were transfected with a miR-30a-5p mimic or miR-30a-5p inhibitor, and the protein levels of Beclin-1 were determined by western blot and semi-quantified. The untransfected cells were used in control group (n=3). (D) mRNA expression levels of Beclin-1 were detected by RT-qPCR. (E) RNA immunoprecipitation assay was performed using anti-Ago2 in HK-2 cells transfected with miR-30a-5p mimic or mimic-nc. miR-30a-5p and 3′UTR of Beclin-1 mRNA expression levels in the anti-Ago2 immunoprecipitated products were measured by RT-qPCR. *P<0.05 vs. miR-30a-5p mimic-nc or control group. miR, microRNA; Mut, mutant; nc, negative control; RT-qPCR, reverse transcription-quantitative PCR; UTR, untranslated region; WT, wild-type.