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. 2019 Aug 23;7(10):e00860. doi: 10.1002/mgg3.860

Figure 4.

Figure 4

MiR‐497‐5p directly targets insulin‐like growth factor 1 (IGF1). (a) The putative miR‐497‐5p binding sites in the 3’UTR of IGF1 mRNA are shown. A mutation was generated on the IGF1 3’UTR sequence in the complementary site for the seed sequence of miR‐497‐5p. (b) Wild‐type (IGF1 3’UTR‐WT) or mutant (IGF 3’UTR‐mut) reporter plasmids were co‐transfected into Huh7 cells with miR‐497‐5p or miR‐GFP. The normalized luciferase activity in the control group was set as the relative luciferase activity. (c) Wild‐type (IGF1 3’UTR‐WT) or mutant (IGF 3’UTR‐mut) reporter plasmids were co‐transfected into HepG2 cells with miR‐497‐5p or miR‐GFP. The normalized luciferase activity in the control group was set as the relative luciferase activity. (d) The mRNA expression of IGF1 was analyzed using RT‐PCR in Huh7 and HepG2. GAPDH was used as an internal control. (e) The protein expression of IGF1 and IGF1R was analyzed using western blotting in Huh7 and HepG2. GAPDH was used as an internal control. All experiments were performed in triplicate with similar results. (f) The correlation between the miR‐497‐5p level and IGF1 mRNA level was measured in the same set of tissues. *p < .05; ***p < .001