Fig. 7: Phosphorylation of RAD51AP1 by NEK2 and CDK1 affects its binding to RAD51.
From: The Greatwall kinase safeguards the genome integrity by affecting the kinome activity in mitosis
a Representative images of kinase assays for wild-type and mutants of full-length murine RAD51AP1 fused to GST and phosphorylated by CDK1/cycB1 complexes or NEK2. 32P Radioactive signal. CB Coomassie blue. b Bar graph of the averaged ratio of radioactive signal of the mutants vs. the wild-type form from three replicates as shown in a. Error bars depict SEM. Phosphatase assay with increasing quantity of immunoprecipitated PP2A/B55 or PP2A/B56 complex in the absence or presence of calyculin A (Cal-A) on prior phosphorylated wild-type full-length murine RAD51AP1 fused to GST phosphorylated by c NEK2 or d CDK1/cycB1. Error bars depict SEM. e Representative blots of co-immunoprecipitation of the RAD51-RAD51AP1 complex transiently expressed in asynchronous or mitotic (M) 293FT cells. Bar graph depicts the signal ratio obtained of the co-immunoprecipitated protein in comparison to the WT asynchronous condition (lane 3).
