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. 2023 Apr 30;35(2):140–162. doi: 10.21147/j.issn.1000-9604.2023.02.05

Figure 5.

Figure 5

SNP rs8400 A/G causes different binding abilities of miR-186-3p to ALKBH5 3’UTR. (A) We found the binding sites for ALKBH5 3’-UTR and miR-186-3p by prediction software (Bartel, 2009); (B) When a miRNA binds to a target, the target first transitions to an intermediate state by unraveling the seed target nucleotides, and then forms a complex upon binding to the miRNA; (C,D) Using RNAfold and RNAcofold WebSever, we found that the rs8400 A allele (D) binds miR-186-3p to a reduced capacity compared with the rs8400 G allele (C); (E) Western blotting results showed that inhibiting miR-186-3p could promote expression of ALKBH5, and increasing miR-186-3p could inhibit the expression of ALKBH5; (F) Luciferase analysis showed that miR-186-3p could bind to the 3’-UTR of ALKBH5 gene, and rs8400 A allele reduced the binding ability of miR-186 to the 3’-UTR. *, P<0.05; **, P<0.01; ***, P<0.001.