Fig. 1

Screening of TbPARP inhibitors. a Inhibition of the in vitro TbPARP (2.5 nM) activity was tested with a library of 31 compounds. Inhibitors were added at 1 μM (grey bars) or 10 μM (white bars). The values were transformed to % of inhibition. Triplicates were measured for every data point, and the mean and the standard deviation are shown. b For the most potent inhibitors, % inhibition was determined at 100 nM concentration. c Inhibition of TbPARP was confirmed in the presence of 1 μM inhibitors by Western blot using 1 μM biotinylated NAD⁺ as a substrate and in the presence of activated DNA 25 μg/mL. TbPARP (120 nM) was incubated in the absence of inhibitors for one hour (positive control) or was added immediately before stopping the reaction (negative control). Synthesized biotinylated PAR was recognized with streptavidin-HRP. Loading of equal amount of protein in every lane was checked with Lysozyme