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. 2019 Mar 29;27(4):459–467. doi: 10.3727/096504018X15193500663936

Figure 3.

Figure 3

miR-664 directly targets the 3′-untranslated region (3′-UTR) of insulin receptor substrate 1 (IRS1) and reduces IRS1 expression in BC cells. (A) Schematic of a putative wild-type (Wt) and mutant (Mut) miR-664 binding site in the 3′-UTR of IRS1. miR-664 mimic or miR-NC was transfected into MCF-7 and MDA-MB-231 cells. At 48 h posttransfection, (B) RT-qPCR analysis was conducted to measure IRS1 mRNA expression. (C) Western blot was used to determine IRS1 protein level at 72 h posttransfection. *p < 0.05 compared with miR-NC. (D) MCF-7 and MDA-MB-231 cells were cotransfected with miR-664 mimic or miR-NC and pMIR-Wt-IRS1-3′-UTR or pMIR-Mut-IRS1-3′-UTR. After 72 h of transfection, luciferase reporter assays were adopted to assess luciferase activity. *p < 0.05 compared with miR-NC.