Supplementary Figure 3: High-affinity binding of S1P and FTY720-P to HDAC1. | Nature Neuroscience

Supplementary Figure 3: High-affinity binding of S1P and FTY720-P to HDAC1.

From: Active, phosphorylated fingolimod inhibits histone deacetylases and facilitates fear extinction memory

Supplementary Figure 3

Recombinant His-tagged HDAC1 immobilized to Ni-NTA-resin was incubated with [32P]S1P (0.1 nM) in the absence or presence of increasing concentrations of unlabeled S1P or FTY720-P. Beads were washed extensively and [32P]S1P bound to HDAC1 was eluted with 500 mM imidazole and radioactivity detected by scintillation counting. Data were fit to a one-site Scatchard model using GraphPad Prism. S1P and FTY720-P bind to HDAC1 with apparent Kd values of 7.5 and 6.2 nM, respectively. R2 values for displacement by S1P and FTY720-P were 0.9581 and 0.9610, respectively.

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