Fig. 2: CDK1 phosphorylates SET isoform 1 at S7 in cells. | Cell Death & Disease

Fig. 2: CDK1 phosphorylates SET isoform 1 at S7 in cells.

From: Cyclin-dependent kinase 1-mediated phosphorylation of SET at serine 7 is essential for its oncogenic activity

Fig. 2

a HeLa cells were treated with DMSO, nocodazole (Noco), or taxol and total cell lysates were probed with the indicated antibodies. Increased cyclin B1 levels mark cells in mitosis. b Control and SET-knockdown (both isoforms 1 and 2) HeLa cells were subjected to the indicated treatment. Total cell lysates were analyzed by western blotting with the indicated antibodies. Increased cyclin B1 levels mark cells in mitosis. c HEK293T cells were transfected with isoform 1 SET or SET-S7A plasmid. At 30 h posttransfection, the cells were treated with nocodazole for 16 h. Total cell lysates were probed with the indicated antibodies. Increased p-Aurora A/B/C levels mark cells in mitosis. d HeLa cells were treated with nocodazole for 16 h and RO3306 (5 µM) or Purvalanol A (10 µM) was added to cells 1.5 h before harvesting as indicated. Proteasome inhibitor MG132 was also added (together with inhibitors) to prevent cyclin B from degradation and cells from exiting from mitosis. Increased cyclin B1 levels mark cells in mitosis. e HEK293T cells were transfected with expression constructs as indicated and total cell lysates were analyzed by western blotting. GFP-Cyc B1-CA: GFP-Cyclin B1-R42A (a nondegradable/constitutive active mutant). Flag-CDK1-CA: Flag-CDK1-T14A/Y15A (nonphosphorylatable/constitutive active CDK1). f HeLa cells were synchronized by a double thymidine (DT) block and release method. Total cell lysates were harvested at the indicated time points and subjected to western blotting analysis. Increased p-Aurora-A and cyclin B1 levels mark cells in mitosis

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