Fig. 1
MARylation of PKCδ by ARTD10 reduces its kinase activity. a In vitro ADP-ribosylation assay of the GST-ARTD10 catalytic domain (818-1025) with His6-PKCδ with or without [32P]-γ-NAD+. BSA was included as substrate negative control. Exposure to X-Ray (32P) was allowed for 96 h. CB, Coomassie blue; 32P, autoradiogram. b Western blot analysis of MARylation of immunoprecipitated PKCδ from HEK293 cells transiently expressing exogenous PKCδ in the presence of ARTD10 or ARTD10-G888W. The expression of the proteins was verified in whole cell lysates (WCL). Treatment with OUL35 or vehicle control (DMSO) was as indicated. c Left, Western blots of phosphorylated PKCδ (Y311), indicating active PKCδ, and of total PKCδ from control HeLa cells or HeLa cells expressing wild type ARTD10 or ARTD10-G888W. Right, ratio of phosphorylated PKCδ/total PKCδ (n = 4). *p < 0.05 (one-way ANOVA followed by Tukey’s test)