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. 2016 Jun 15;9:3535–3544. doi: 10.2147/OTT.S106351

Figure 3.

Figure 3

miR-503 directly targets IGF-1R.

Notes: (A, B) TargetScan software predicted that IGF-1R was a target of miR-503, and this targeting relationship was evolutionally conserved. (C) The seed sequences of miR-503 in the wild type (WT) or mutant type (MUT) of IGF-1R 3′-UTR are indicated. (D) The WT or MUT IGF-1R 3′UTR were cloned into luciferase reporter vector. (E) The luciferase activity was significantly decreased in 293T cells cotransfected with the WT IGF-1R vector and miR-503 mimic, but unaltered in 293T cells cotransfected with the MUT IGF-1R vector and miR-503 mimics, compared to the control group. Control, 293T cells transfected with only WT IGF-1R or MUT IGF-1R vectors; NC, 293T cells cotransfected with WT IGF-1R or MUT IGF-1R vector and scramble miR. (F) Real-time RT-PCR was performed to determine the miR-503 levels in LM3 and HepG2 cells transfected with miR-503 mimic and miR-503 inhibitor, and (G) the protein levels of IGF-1R were then examined using Western blot assay. Nontransfected cells were used as control. **P<0.01 vs control. ***P<0.001.

Abbreviations: RT-PCR, reverse-transcription polymerase chain reaction; miR, microRNA; IGF-1R, insulin-like growth factor 1 receptor; UTR, untranslated region; CMV, cytomegalovirus; PCT, probability of conserved targeting.