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. 2009 Jul 13;122(15):2750–2759. doi: 10.1242/jcs.048223

Fig. 4.

Fig. 4.

The kinase activity of Plk1 is required for the interaction between Plk1 and FAM29A, but not for the interaction between FAM29A and NEDD1 or between Plk1 and NEDD1. (A) HeLa S3 cells were synchronized at prometaphase by a thymidine-nocodazole arrest, released and incubated for 2 hours with medium containing the proteasome inhibitor MG132 (20 μM) and one of the followings: DMSO (lane 2), purvalanol A (25 μM; lane 3), BI 2536 (1 μM; lane 4), or purvalanol A (25 μM) + BI 2536 (1 μM; lane 5). Asynchronous cells without drug treatment (lane 1) were also included. Cell lysates and the anti-FAM29A and anti-Plk1 immunoprecipitates (IP) were analyzed by western blotting. Arrowheads point to nonspecific cross-reacting proteins. (B) HeLa cells were transfected with siRNAs and synchronized at prometaphase by a thymidine-nocodazole treatment. The anti-FAM29A immunoprecipitates (IP) were analyzed by western blotting.