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. 2019 Jul 18;21(9):849–862. doi: 10.1016/j.neo.2019.06.004

Figure 2.

Figure 2

miR-125a-5p modulates cell proliferation in vitro via regulating the cell cycle at the G1/S transition by increasing p27 expression. (A) Transfection of miR-125a-5p into HN5 and UM-SCC-22B cell lines results in decreased cell counts, compared to transfection of a control miRNA. (B) Transfection of miR-125a-5p into HN5 and UM-SCC-22B cell lines results in luminenscence, using the CellTiter-Glo Assay (Promega), compared to transfection of a control miRNA. (C) Transfection of miR-125a-5p into HN5 and UM-SCC-22B cell lines results in increased proportion of cells in the G1 phase with a concomitant decrease in percentage of cells in S phase, compared to control miRNA transfection, as determined by the propidium iodide cell cycle assay. Cell cycle analysis was performed on the FlowSight Imaging Flow Cytometer (Amnis). (D) Western blotting of p27 protein shows increased expression of p27 after transfection of miR-125a-5p mimic, as compared to control miRNA. 5-bromo-2′-deoxyuridine (BrdU) assay reveals that miR-125a-5p does not affect DNA synthesis in HN5 (E) and UM-SCC-22B (F) cell lines. Data were analyzed with the Student's t-test and are presented as the mean ± standard deviation. Experiment was carried out in triplicate with the BrdU DNA synthesis assay performed in quintuplicate.