Figure 2.

Tet2 generates fC and caC early and without a requirement for hmC accumulation. (A) Example traces for [¹⁴C]-mC Tet activity assay. DNA duplexes from Tet reactions were degraded and spiked with standards to delineate fractions containing each base. The fractions were then subjected to scintillation counting. Top: Chromatogram of nucleoside standards (10 μL of 10 μM each). Bottom: Corresponding LSC trace. (B) Time course of Tet2 (10 μg/mL) turnover of 500 nM [¹⁴C]-mCpG/CpG duplexed DNA showing total activity and fractions of each ox-mC at 1 and 20 min. (C) Titration of 75–4000 nM [¹⁴C]-mCpG/CpG with 5 μg/mL Tet2, reacted for 10 min. Total specific activity is plotted on the left y-axis (black bars) as mean and SD of duplicate experiments, along with fraction of each ox-mC base on the right y-axis.