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Figure 5. Aurora A phosphorylated TACC3 is enriched at mitotic centrosomes. (A) Immunolocalization of TACC3 and phospho-TACC3. Deconvolved images of human tissue culture cells stained for DNA (blue)/microtubules (MTs; green), TACC3, and phospho-TACC3 (P-TACC3; stained by antiphospho-Ser626 antibodies) in different cell cycle stages. Bars, 10 μm. (B) Immunolocalization of TACC3 and phospho-TACC3 in siRNA-treated cells. Deconvolved images of either control or TACC3 RNA interference–treated cells stained for DNA (blue)/microtubules (MTs; green), TACC3 (top), and phospho-TACC3 (P-TACC3; bottom). Arrows indicate misaligned chromosomes that are indicative of the TACC3 RNA interference phenotype. Bar, 10 μm. (C) Phosphorylation of TACC3 is mitosis specific in human tissue culture cells. The TACC3 antibody detects a protein of ∼130 kD in extracts prepared from either an asynchronous culture of HeLa S3 cells (lane 1; I) or cells arrested in mitosis by nocodazole treatment (lane 2; M). The phosphospecific TACC3 antibody (antiphospho-Ser626) recognizes a band with the same molecular mass size in mitotic cells (lane 4) but not in asynchronously cultured cells (lane 3). The blot probed by α-tubulin antibody is a loading control (lane 5, asynchronous cultured cells; lane 6, mitotic arrested cells). Image published in: Kinoshita K et al. (2005) Copyright © 2005, The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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