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Mutagenic analysis of crucial WWE domain residues involved in iso-ADPR binding. (A) Dissociation constants of RNF146 WWE domain mutants with iso-ADPR as measured by ITC analysis. (B) Coimmunoprecipitation of wild-type RNF146—but not RNF146 mutants Y107A, Y144A, Q153A, and R163A—by anti-PAR antibody in HEK293 cells. (C) Expression of siRNA-resistant wild-type RNF146—but not RNF146 mutants Y107A, Y144A, and Q153A—rescues RNF146 siRNA-induced stabilization of Axin1 in HEK293 cells.
