Figure 1.
Fbxo6 is phosphorylated during mitosis. (a-d) HEK293 cells were transfected with flag-tagged Fbxo6 (a), Fbxo4 (b), Fbxo22 (c) or Fbxo36 (d) for 24 hours, respectively. Then the cells were treated with nocodazole (200 ng/ml) for 16 hours (Add for addition) and released for 8 hours (Rel for release). The cells were collected and equal amounts of cell lysates were blotted for anti-flag antibody. (e) HeLaflag-Fbxo6 cells were treated with or without nocodazole (200 ng/ml) for 16 hours. Cell lysates were collected in the presence or absence of lambda protein phosphatase (λ-ppase) and the indicated proteins were examined by western blot. (f) HeLaflag-Fbxo6 cells were treated with nocodazole (200 ng/ml) for 16 hours, and then with or without MG132 (20 μM) for another 3 hours. At the meantime, cells were treated with Roscovitine (20 μM) or Volasertib (50 nM) for 3 hours as well, and the indicated proteins were examined by western blot.