Figure 1.
Generation of Partially Reprogrammed iPSCs by SeVdp Vectors
(A) Structures of SeVdp(KOSM) and SeVdp(GKOSM).
(B) Expression of ESC markers. Colonies induced by SeVdp(KOSM) or SeVdp(GKOSM) were stained for ALP or SSEA-1 at day 10 or 18, respectively. Scale bars, 100 μm.
(C) Time course of mRNA expression. Cells were collected at day 0, 6, 13, 25, and 49, and the mRNA levels of indicated genes were determined by quantitative RT-PCR and plotted relative to those in MEFs or mouse iPSCs (miPSC) (Nishimura et al., 2011). Data represent means ± SEM of three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 versus SeVdp(KOSM)-infected cells.
(D) Reproducibility of reprogramming by SeVdp(GKOSM). Cells were collected at day 25 (Exp. 1) or day 23 (Exp. 2 and 3), and the mRNA levels of Cdh1, Fgf4, and Rex1 were determined by quantitative RT-PCR. Data represent means ± SEM of three independent PCR reactions. n.s., not significant; ∗∗∗p < 0.001 versus Exp. 1.
(E) Protein levels of reprogramming factors expressed from SeVdp vectors. SeVdp(KOSM) or SeVdp(GKOSM)-infected MEFs were collected 1 day after infection, and the protein level of each reprogramming factor was determined by western blotting as described in the Experimental Procedures. The protein level of each factor is plotted relative to that in SeVdp(KOSM)-infected cells. Data represent means ± SEM of three independent experiments. ∗∗p < 0.01.