Figure 3.

TET1/2-Mediated 5hmC Modification Plays Important Roles in Rapid iPSC Induction by OySyNyK Factors

(A) qRT-PCR assays showing the expression levels of Tet1, Tet2, and Tet3 in OSNK- and OySyNyK-transduced MEFs at different time points. Both iPSCs and ESCs were included for the comparison. OSNK- and OySyNyK-transduced MEFs groups were compared to MEF groups. ^∗∗∗p < 0.001 versus control groups.

(B) Dot-blot assays showing the 5hmC level in OSNK- and OySyNyK-transduced MEFs at different time points. ESCs (CGR8), iPSCs, and MEFs cells were used as controls.

(C) Immunostaining of 5hmC in MEFs and OSNK- and OySyNyK-transduced MEFs at day 2.

(D) Immunostaining assays showing that OySyNyK-transduced GFP-positive MEFs contained high levels of 5hmC at day 2.

(E) qRT-PCR assays showing the efficiencies of Tet1 and Tet2 knockdown in OySyNyK-transduced MEFs. ^∗∗∗p < 0.001 versus control groups.

(F) Statistical summary showing the number of GFP-positive colonies induced by OySyNyK under Tet1 or Tet2 knockdown. ^∗∗∗p < 0.001 versus control groups.

(G) Alkaline phosphatase (AP) staining showing the efficiencies of iPSC colony induction by OySyNyK under Tet1 or Tet2 knockdown.

Data (A), (E), and (F) are from at least three independent experiments (mean and SD of triplicate assays). Groups were compared using the Student’s t test. ^∗∗∗p < 0.001 versus control groups.