File:Cellular-Contractility-Requires-Ubiquitin-Mediated-Proteolysis-pone.0006155.s010.ogv

English: Nocodazole induced contractions depend on neddylation. E36-wt or E36-ts41 cells were treated with nocodazole at the permissive and the restrictive temperatures. A, E36-wt control cells undergo normal mitosis at 39.5°C. B, An E36-wt cell grown at 39.5°C was treated with nocodazole and contracted upon entry into mitosis. C, An E36-ts41 cell grown at 34°C was treated with nocodazole. The cell arrested and contracted for 4 hours before it flattened. D, E6-ts41 cells undergo cell cycle arrest at 39.5°C. The movie started at 34°C and the cell (arrow, 44 minutes) underwent normal mitosis. Fifteen hours later the temperature was shifted to 39.5°C for an additional 11 hours. The cell was overall arrested for at least 26 hours. The cell cycle time of these cells at 34°C is about 16 hours. E, Temperature shift to 39.5°C of nocodazole-treated, mitotic E36-ts41 cells stopped their contractions. At the beginning of the movie the temperature was shifted to 39.5°C and nocodazole was added. Upon mitotic entry the cell (arrow, 52 minutes) contracted for 14.5 hours (944 minutes), and then stopped for an additional 4 hours. F, Detached control E36-wt (left panel) and E36-ts41 (right panel) were allowed to re-adhere on a glass cover slip. Nocodazole was added and the temperature was shifted to 39.5°C at the beginning of the movie. While control cells contracted and re-adhered within 4–8 hours, E36-ts41 cells contracted for 10 hours and then ceased. These cells failed to adhere the matrix.