Skip to main content
. Author manuscript; available in PMC: 2021 Jul 6.
Published in final edited form as: Angew Chem Int Ed Engl. 2020 May 11;59(28):11312–11315. doi: 10.1002/anie.202002751

Figure 3.

Figure 3.

TET2 direct N-demethylation activity is comparable to 5mC oxidation activity. (A) Product formation can be detected by either protection from MspI cleavage, for F27–5mC, or susceptibility to cleavage for F27–4dmC. Fixed concentrations of F27–4dmC, F27–5mC and F27–5mZ substrates (200 nM) were co-incubated with serial dilutions of TET2-CS (6 nM to 1.6 μM), then subjected to the appropriate MspI-based assay. Representative product gels, imaged for FAM fluorescence, are shown. (B) Percent conversion of F27–5mC, F27–5dmC, and F27–5mZ are shown as a function of TET2-CS concentration. Error bars represent standard deviation from three independent replicates and the best fit sigmoidal curve to aggregated data is shown. The EC50 value derived from this fit represents the amount of enzyme required to convert half of the substrate, with 95% confidence interval (CI) noted.