Nuclear proteins from iHepSCs and MEFs were prepared with a Nuclear Extraction Kit (ab113474, Abcam) and quantitated with Optiblot Bradford Reagent (ab119216, Abcam). The activity of the Tet family of 5mC hydroxylases, including TET1, TET2 and TET3, was detected by a TET Hydroxylase Activity Quantification kit (Colorimetric) (ab156912, Abcam) according to the manufacturer’s instructions. The amount of nuclear extracts for each assay was 10 µg per sample. The optical value was measured at 450 nm with an optional reference wavelength of 655 nm by Molecular Devices Spectra MAX190. The activity of the Tet enzyme was calculated as follows: Tet activity (OD/min/mg) = (sample OD – blank OD)/(protein amount (µg) × 90(min)) × 1000.
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