Figure 6
miR-302 contributes to the pluripotency and tumorigenicity in hPSCs by maintaining OCT4 at high expression level through suppressing of AKT1. (a) The expression of miR-302s was analyzed during the process of EB formation at days 5, 10, 15, 20, 25 and 30 by TaqMan qRT-PCR assays. U6 was used as an internal normalization control. Data are presented as mean±S.D. (n=3) and are representative of three independent experiments. (b) The expression of AKT1, OCT4 and SOX2 were assessed by western blot during the differentiation of hESCs and hNT-2 cells to EBs. GAPDH was used as a loading control. (c) The western blot analysis evaluated the expression of AKT1 and OCT4 when miR-302 was upregulated by transfected miR-302s mimics in AKT1 overexpressing hNT-2 cells. A negative control oligonucleotide was transfected as control, and GAPDH was used as a loading control. (d) Western blot analysis was used to evaluate the expression of AKT1 and OCT4. A negative control oligonucleotide antagomir was transfected as a control, and GAPDH was used as a loading control. (e) AKT1 overexpressing vectors were transfected into hNT-2 cells for 5 days, and the morphological changes of AKT1-overexpressed hNT-2 cells and negative control cells were observed under light microscope. Scale bars, 100 μm
