Fig. 5.
Assay of MTHFR activity following addition of AdoMet, (S)-SKI-72 or sinefungin. A-C. Remaining activity of hsMTHFRFL following incubation with increasing concentrations of AdoMet (A), (S)-SKI-72 (B) or sinefungin (C). D. Remaining activity of hsMTHFRFL with increasing concentrations of AdoMet following pre-incubation with 100 μM sinefungin. E-F. Remaining activity of endogenous MTHFR from HEK293 cell lysate following incubation with increasing concentrations of AdoMet (E) or (S)-SKI-72 (F). G. Remaining activity of endogenous MTHFR from HEK293 cell lysate following incubation of (S)-SKI-73 with live cells for 12 or 48 h. Repl: replicate. Each curve represents a single technical replicate, of which n = 3 were performed in each assay. Curve was fit and inhibitor constant (Ki) calculated as described in the Materials and Methods.