Figure 3
miR-302 regulates cell cycle progression by promoting the G1 to S transition. (a) Flow cytometry analysis of the cell cycle distribution in hMSCs when exogenous expression of miR-302 was upregulated. A lentiviral vector expressing a scrambled sequence was used as a control. (b) Flow cytometry analysis of the cell cycle distribution in hESCs and hNT-2 cells when endogenous expression of miR-302s was suppressed by miR-302s antagomir. Negative control oligonucleotide antagomir was transfected as control. (c) Western blot analysis in hESCs and hMSCs of the expression levels of key regulators involved in the G1 to S transition; GAPDH was used as a loading control. (d) Immunofluorescence detected the expression of proteins by Hoechst 33342 counterstaining. Scale bars, 200 μm. (e) qRT-PCR measured the mRNA expression of key regulators involved in the G1 to S transition. GAPDH was used as an internal normalization control. Data are presented as mean±S.D. (n=3) and are representative of three independent experiments. (f) Targetscan, PicTar and Miranda predicated the cell cycle-associated candidate targets of miR-302. Green indicated positive regulators, red indicated negative regulators
