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. Author manuscript; available in PMC: 2008 Apr 30.
Published in final edited form as: Dev Cell. 2007 May;12(5):699–712. doi: 10.1016/j.devcel.2007.03.014

Fig. 5. Phosphorylation of GEF-H1 by mitotic kinases Aurora A and Cdk1/Cyclin B.

Fig. 5

Inhibition of Aurora kinases and Cdk1/Cyclin B abolish GEF-H1 phosphorylation during mitosis. Before adherence, harvested synchronized HeLa cells were inhibitor treated for 15 min during release into fresh culture medium (w/o nocodazole) supplemented with the same inhibitors (at denoted concentrations and specificities). Inhibitors used here are specific for serine-threonine kinases that are either predicted by the Scansite algorithm (scansite.mit.edu) to phosphorylate GEF-H1 (PKA, GSK3β; data not shown) or are required for the control and timing of mitotic progression (Aurora A/B, Cdk1/Cyclin B) (Ferrari, 2006; Nigg, 2001). At the indicated times (after adherence), cells were lysed and analyzed by anti-pGEF-H1 and anti-GEF-H1 immunoblotting.