Figure 3.

CDK16 directly interacts with p53 in vivo and in vitro. A. HEK293T cells transfected with the indicated constructs were collected 24 h later. Cells were lysed with NETN buffer. Immunoprecipitation (IP) using S-protein agarose were performed and Western blotted with the indicated antibodies. WCL: whole cell lysate. B. Endogenous CDK16 associated with p53 in A549 cells. A549 cells were lysed and subjected to immunoprecipitation using anti-IgG or anti-p53 as indicated, and were analyzed by Western blotting using indicated antibodies. C. Beads coated with GST or GST-p53 fusion proteins were incubated with SFB-CDK16 protein overnight. GST pulldown was immunoblotted with indicated antibodies. D. Schematic description of p53 domains and deletion mutants used in this study. E. HEK293T cells co-transfected with constructs encoding SFB-CDK16 and indicated p53 mutants were collected 24 h later. Cells were lysed, the supernatants were incubated with S-protein agarose beads, and then analyzed by Western blotting using antibodies as indicated.