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. 2019 Nov 27;12:10311–10321. doi: 10.2147/OTT.S190998

Figure 3.

Figure 3

miR-99a targeting the mTOR gene in CRC cells. HCT-116 cells were transiently transfected with 100 nM miR-99a mimic or negative control (NC) for 48 h. The relative mTOR mRNA expression levels were determined by qRT-PCR (A). The mTOR protein expression levels were determined by Western blotting (B). The putative mir-99a binding site in mTOR 3′-UTR and an alteration of the mTOR 3′-UTR bearing mutant in the seed sequence are shown. The dual luciferase assay was performed to confirm the direct regulation of miR-99a on mTOR 3′-UTR (C). The relative mTOR mRNA expression levels were determined by qRT-PCR in the HCT-116 cells transfected with miR-99a mimic or NC (D). The mTOR protein levels were determined by western blotting in the HCT-116 cells transfected with miR-99a mimic or NC (E). (**P<0.01, ***P<0.001).