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. 2016 Aug 13;7(38):61166–61182. doi: 10.18632/oncotarget.11278

Figure 7. MiR-4653-3p downregulated FRS2 expression by binding to two complimentary sites on 3′UTR of FRS2 mRNA.

Figure 7

(A) Two sites located in 84~90 and 2213~2219 on 3′ UTR of FRS2 mRNA were predicted to be complimentary to miR-4653-3p by multiple databases, miRDB, TargetScan and DIANA. (B) 293T cells were transfected with control mimics or miR-4653-3p mimics together with the pmirGLO Vector constructs contained a predicted binding sequence (FRS2 84 or FRS2 2213). Forty-eight hours after transfection, cells were analyzed for luciferase activity using the Dual-Glo® Luciferase Assay System. The bars represent the mean ± standard deviation of at least 3 independent experiments for each condition. * indicates significant decrease of normalized firefly luciferase activity compared to controls and control mimics. P < 0.0001 as calculated by One-way ANOVA and LSD test. (C) Real-time RT-PCR results for miR-4653-3p level was showed. ** indicates significant overexpression of miR-4653-3p compared to control and pGLV3-NC. P < 0.001 as calculated by One-way ANOVA and LSD test. Control, indicates untransfected cells. (D) MCF7-TAMR and BT474-TAMR cells were infected with lentivirus particles which mediate miR-4653-3p expression (pGLV3-miR-4653) or the negative control (pGLV3-NC). Western blot results for FRS2 protein were showed.