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. 2020 Sep;9(9):5190–5199. doi: 10.21037/tcr-20-1083

Figure 4.

Figure 4

hsa-miR-195-5p suppresses cyclin D1 expression by directly targeting the cyclin D1 3'-UTR in TPC-1 cells. (A) TargetScan prediction of hsa-miR-195-5p targeting of cyclin D1 and Mutagenesis of cyclin D1 3'-UTR. (B) Luciferase reporter assay. (C) Effects of hsa-miR-195-5p mimics on the endogenous cyclin D1 mRNA levels. TPC-1 cells were cotransfected with hsa-miR-195-5p mimics or negative control oligonucleotides. Forty-eight hours after transfection, cells were isolated, the expression of cyclin D1 was analyzed by qRT-PCR. (D,E) Effects of hsa-miR-195-5p on the endogenous cyclin D1 protein levels. Forty-eight hours after transfection with the hsa-miR-195-5p mimics or negative control oligonucleotides, proteins were isolated from TPC-1 cells and analyzed by Western blotting, with GAPDH as an internal control. ****, P< 0.0001 compared with respective controls. NC, negative control oligonucleotides.