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Lysate from J774A.1 cells (20 µg) was either left untreated for control (con) or incubated for 10 min at 37°C with either C3bot1E174Q-C2I (0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 10 ng/mL) or C2I (0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 10 ng/mL) in the presence of biotin-NAD+ (10 µM). Samples were subjected to SDS-PAGE, blotted and biotinylated (i.e. ADP-ribosylated) actin was detected with streptavidin-peroxidase.
