β-TrCP-Mediated Degradation of Cyclin F Promotes Mitotic Progression
(A) HeLa cells stably expressing an empty vector (EV), cyclin F WT, and SS > AA 700/704 were synchronized by a DTB. Cells were collected at the indicated hours after release, lysed, and immunoblotted as indicated.
(B) HeLa cells stably expressing an empty vector (EV), cyclin F WT, and SS > AA 700/704 were synchronized as in (A). Extracts were collected at the indicated hours, immunoprecipitated (IP) with anti-FLAG resin, and immunoblotted as indicated.
(C) Quantitative real-time PCR analysis of PLK1 (left panel) and Aurora B (right panel) mRNA levels in HeLa cells stably expressing the indicated plasmids. Student’s t test. ∗p < 0.05, ∗∗p < 0.01.
(D) HeLa cyclin F knockout (KO) cells infected with retroviruses expressing the indicated plasmids were immunoblotted as indicated. The asterisk corresponds to an unspecific band detected by the cyclin F antibody.
(E) HeLa cyclin F knockout (KO) cells stably expressing cyclin F WT and SS > AA 700/704 were synchronized as in (A). Cells were collected at the indicated hours, lysed, and immunoblotted as indicated.
(F) HeLa cyclin F knockout (KO) cells stably expressing cyclin F WT and SS > AA 700/704 were synchronized as in (A). Time-lapse microscopy was performed for 16 hr after release and analyzed. Student’s t test. ∗p < 0.05.
(G) Scheme outlining the mechanism of cyclin F recognition by β-TrCP and its relevance.