Mfn 1 interacts with Cdk1/cyclin B1. (A) G2/M-phase cells were obtained by harvesting cells after nocodazole treatment. Lysates were immunoprecipitated with an anti-cyclin B1 antibody, followed by immunoblotting with the indicated antibodies. (B) G2/M-phase cells were obtained by taxol treatment for 12 h. The association to cyclin B1 was analyzed by immunoblotting, after lysates were immunoprecipitated with an anti-Mfn1 antibody. (C) Cells were treated with MG132 with or without nocodazole. Lysates were immunoprecipitated with an anti-cyclin B1 antibody, followed by immunoblotting with the indicated antibodies. Arrows (NS, upper two bands) indicate non-specific binding. (D) Cells were synchronized at G1/S, G2 or mitotic phase by treatment with thymidine, roscovitine (Ros, 100 μM) for 5 h at 7 h after DTB release or nocodazole for 2 h at R8 after DTB release, respectively. Mitochondrial morphology was observed by staining with MitoTracker Red. Images were analyzed by confocal microcopy. Scale bars = 10 μm.