FIGURE 1.

PHF8 interacts with cyclin E-CDK2. A, coimmunoprecipitation assay. 293T cells were transfected with myc-tagged cyclin E and CDK2 and FLAG-tagged PHF8. Cell lysates were harvested for immunoprecipitation (IP) with FLAG antibody, followed by immunoblotting (IB) with myc antibody. TCE, total cell extract. B, 293T cell lysates were immunoprecipitated with CDK2 antibody or normal mouse IgG and then immunoblotted with PHF8 antibody. C–E, GST, GST-PHF8, or GST-PHF8 mutants, as indicated, immobilized on glutathione beads were incubated with His-CDK2 in lysis buffer for 2 h. The associated protein was eluted with SDS loading buffer and immunoblotted with CDK2 antibody. The asterisks represent GST or GST fusion proteins. CBB, Coomassie Brilliant Blue. NMS, normal mouse serum.