Expression of wild-type or mutant PC-2 does not affect proliferation or STAT-1/p21/Cdk2 activity in NRK-52E cells. (A). Whole cell lysates containing equal amounts of protein from NRK-52E cells trasnsiently transfected with vector-only, HA-WT PKD2, HA-R742X PKD2 and HA-1–702 PKD2 were analyzed by Western blotting for expression of p21, phosphorylated STAT-1, PCNA, tubulin, HA and PC-2. A non-specific band is detected in vector-only and WT PKD2 lanes in the HA blot and co-migrates with mutated PC-2 in this cell line. (B) Cdk2 immunoprecipitates from each transfectant were subjected into an in vitro Cdk2 kinase assay using Histone 1A as substrate. Equal amount of Cdk2 was confirmed by immunoblotting the precipitates with anti-Cdk2 antibody. Data are representative of three independent experiments performed. No statistically significant difference was detected.