Application of ProtoArrays® to identify ARTD10 and ARTD8 substrates. A. Indicated proteins were spotted on nitrocellulose membranes and subsequently incubated with TAP-ARTD10 and biotin-labeled NAD+ to optimize array conditions. The signals were detected with HRP-coupled streptavidin. B. Schematic representation of ProtoArray®-based screens. The arrays were incubated with ARTD10, ARTD8 or BSA and 25 μM biotin-β-NAD+ and visualized using streptavidin-AlexaFluor® 647. C. An exemplary magnification of a sub-array from an array incubated with TAP-ARTD10 is shown with controls and top-outlier PDGF-B indicated. D. Venn diagram showing the overlap between the identified ARTD8 and ARTD10 substrates. E. All putative ARTD8 and ARTD10 substrates sorted according to function.