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. 2018 Jul 30;24:5282–5291. doi: 10.12659/MSM.910163

Figure 5.

Figure 5

MiR-646 inversely regulated IGF-1 mRNA and protein expression. The mRNA expression of IGF-1 was upregulated with miR-646 downregulation and downregulated with miR-646 overexpression in (A) 21% O2 condition (* P=0.03 pre-miR-646 vs. pre-miR-Ctrl, * P=0.02 anti-miR-646 vs. anti-miR-Ctrl); (B) 5% O2 condition (* P=0.03 Pre-miR-646 vs. pre-miR-Ctrl, * P=0.02 anti-miR-646 vs. anti-miR-Ctrl); and (C) 1% O2 condition (* P=0.02 pre-miR-646 vs. pre-miR-Ctrl, * P=0.02 anti-miR 646 vs. anti-miR-Ctrl). The results of western blot showed that miR-646 significantly repressed IGF-1 protein expression in (D) 21% O2 condition; (E) 5% O2 condition; and (F) 1% O2 condition. (G) The direct binding between miR-646 and 3′-UTR of IGF-1 was detected by luciferase activity assay. The wild-type or mutated 3′-UTR of IGF-1 containing binding sites in luciferase reporters was transfected into hPDLCs with miR-646 overexpression or negative control (* P=0.02 pre-miR-646 vs. pre-miR-Ctrl). (H) The predicted miR-646-binding site in the IGF-1 3′-UTR.