Overexpression of BTAK/Aurora-A in ovarian carcinoma.

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Overexpression of BTAK/Aurora-A in ovarian carcinoma. Overexpression of BTAK/Aurora-A in ovarian carcinoma. A, Western blotting analyses of human primary ovarian tumor specimens (T) and normal ovarian tissue (N). Sixty μg of protein from each specimen were separated in SDS-PAGE, transferred, and detected with anti-BTAK/Aurora-A antibody (top). A weak low molecular weight band detected in N, T16, and T17 could be an alternatively spliced form because preincubation of BTAK/Aurora-A antibody with GST-BTAK/Aurora-A antigen is able to compete this band (data not shown). Equal loadings were indicated by hybridization of the same blot with anti-β-actin antibody (bottom). B, Northern blotting analyses of expression of BTAK/Aurora-A mRNA with [32P]-dCTP-labeled BTAK/Aurora-A cDNA as probe (top). The quality and loading amount of RNA were indicated by 28S and 18S ribosome RNA (bottom). In C, Southern blots containing 10 μg of PstΙ-digested DNA per lane from primary human ovarian tumors and a normal ovary control were hybridized with a BTAK/Aurora-A cDNA probe. The location of the 2.8 and 2.5 kb of BTAK/Aurora-A fragments is marked. Blot was stripped and reprobed with a human β-actin probe to confirm equal loading. Tatiana M. Gritsko et al. Clin Cancer Res 2003;9:1420-1426 ©2003 by American Association for Cancer Research