A, Whole-cell lysates from ELT-3 cells were immunoprecipitated using a control antibody or an anti-p27 antibody. The immunoprecipitated pellets were then resuspended and immunoblotted with anti-p27 and anti-14-3-3 antibodies. B, HEK293 cells, which were transfected with Flag-p27-WT, were treated as indicated and used to generate whole-cell lysates. Lysates were analyzed for exogenous p27 expression or utilized for affinity purification using GST-14-3-3 or GST alone and analyzed by immunoblotting with an anti-p27 antibody. In addition, lysates were used for immunoprecipitation with an anti-p27 antibody, and immunoprecipitates were analyzed by immunoblotting using an anti-14-3-3 antibody.