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CK1α phosphorylates NSP2 on S313. NSP2 was detected in a reducing gel by WB with anti-dNSP2 monoclonal antibody. The blot was exposed to autoradiography film overnight to detect the transfer of radiolabeled phosphate to NSP2. CK1α phosphorylation of NSP2 was determined using rNSP2 double mutants H225A S313D and H225A S313A. The autophosphorylation-deficient H225A single mutant, that is only phosphorylated by CK1α, was used as a positive control.
