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. 2011 Jun 8;2(8):592–596. doi: 10.1021/ml2000824

Figure 5.

Figure 5

“In cell” dose-dependent inhibition of human DDAH-1. (A) HEK 293T cells treated with the DDAH activity probe N-but-3-ynyl-2-chloroacetamidine (ActPrb, 154 μM) and increasing ebselen concentrations are washed, lysed, derivatized with azide-labeled biotin, and analyzed by a two color Western blot to show equal loading of DDAH (red) and a decreasing number of active sites available for biotinylation by the activity probe (yellow). The left-most lane contains molecular weight markers (also in red). (B) Dependence of relative fluorescence of biotin-tagged DDAH on ebselen concentration gives an IC50 value of 36 ± 7 μM and Hill coefficient of 0.9 ± 0.2.