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Fig. 2. (A) Purification and characterization of recombinant hPARP-3 overexpressed in the Sf9/baculovirus system. Crude extract from infected Sf9 cells (lane a); Purified recombinant hPARP-3 (lanes b-e); DNA-binding activity of hPARP-3 detected by south-western blotting (lane c); autopoly(ADP-ribosyl)ation of purified hPARP-3 incubated with [{alpha}-32P] NAD+ (lane d); inhibition of hPARP-3 autopoly(ADP-ribosyl)ation by 2 mM 3-Aminobenzamide (lane e). (B) Western blot detection of hPARP-3 in crude extracts from mouse lung (lane f), HeLa cells (lane g) or infected Sf9 cells (lane h) and purified recombinant hPARP-3 (lanes i,j) using two different anti hPARP-3 antibodies (see Materials and Methods).





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