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. 2015 Mar 23;112(14):4316–4321. doi: 10.1073/pnas.1417939112

Fig. 1.

Fig. 1.

Enzymatic activity of NgTET1 on oligo, plasmid and gDNA. (A) RE-based assay showing protection of NgTET1-treated pRS(M.HpaII) plasmid against digestion with MspI at varying concentrations of NgTET1. (B) LC-MS (Agilent 1200)-based assay reflecting NgTET1 reaction species using mammalian gDNA IMR90. Reactions contained 1.5 μg sheared (1.5-kb) DNA and 4 μM NgTET1. (C–E) Quantification of NgTET1 reaction species as measured by LC-MS (Agilent 1200 for C and D; 6490 Triple Quad LC-MS for E) for different types of DNA. The error bars (in black) represent the SEM (n ≥ 3). (C) Two micromolars oligo (Table S1), 1.5 μg plasmid, and 1.5 μg gDNA were used with 4 µM NgTET1. (D) Four micromolars 5mC sites for symmDNA, hemiDNA and ssDNA (Table S1) were used with 8 μM NgTET1. (E) Two micromolars ds- or ss-oligo (Table S1) or sheared (1.5-kb) HeLa (0.5 μg) or M.Fnu4HI (0.2 μg) gDNA were used with 6.7 µM NgTET1 (in Mops buffer pH 6.9) or mTET1CD.