The rapid G/M transition of HeLa cells over-expressing TSPY is associated with an early degradation of the mitotic cyclin B1

Copyright information:Taken from "TSPY potentiates cell proliferation and tumorigenesis by promoting cell cycle progression in HeLa and NIH3T3 cells"BMC Cancer 2006;6():154-154.Published online 9 Jun 2006PMCID:PMC1526451. HeLa Tet-off cells harboring the vector alone (A) and TIG-TSPY construct (B) were synchronized at G/S, released into S phase, harvested and analyzed with western blotting with various antibodies at 2-hour intervals for 22 hours. The mitotic cyclin B1 is synthesized normally in late S phase and Gand is rapidly degraded before the cell exits mitosis. In cells over-expressing EGFP in the vector alone, the levels of cyclin B1 increased at 6 hours after the G/S release, peaked at 10 hours and gradually decreased until 14–16 hours (A, top row) while in those over-expressing both EFGP and TSPY, the levels of cyclin B1 increased gradually at 2 hours after the G11/S release, peaked at 10 hours and rapidly reduced thereafter (B, top row). The filters were re-probed with tubulin antibody (A, bottom row; B, middle row) showing relatively even loading of the samples. TSPY was uniformly expressed and detected in samples of cells over-expressing this transgene (B, bottom row).