Identification of GIG47 and its expression patterns in human breast cancers, multiple human tissues and cancer cell lines. (A) Comparative gene expression profile examined by DDRT-PCR using total RNA isolated from normal breast tissue, breast cancer tissue, and MCF-7 breast cancer cell line. Differential display was carried out 5' arbitrary primer H-AP21 (5’-AAGCTTTCTCTGG-3’) and 3’ H-T11C anchored primer (5’-AAGCTTTTTTTTTTTC-3’; GenHunter). The PCR products were resolved by electrophoresis. HP73 is the name of the partial GIG47 gene product. The arrow identifies the location relative to other PCR products. (B) The expression level of GIG47 was examined by the quantitative PCR analysis in various breast cancer cell lines compared to the normal breast tissues. (C) Total RNAs were isolated from normal breast tissues and their corresponding primary breast cancer tissues. Blot was hybridized with the randomly primed [32P]-labeled GIG47 partial cDNA probe (the HP73 fragment). Human β-actin cDNA was used as a control probe (lower panel). Northern blotting was performed to determine the expression of GIG47 in different human tissues. Normal 12 lane multiple tissue northern blot (D) or human cancer cell line multiple northern-blot (E) purchased from Clontech were probed with a radioactively labeled HP73 partial cDNA (upper panel) or human β-actin cDNA control probe provided by Clontech (lower panel). (E) The normal cell lines included here were BJ, IMR-90, and MCF12F which are a human fibroblast from normal foreskin, a human lung cell fibroblast, and an epithelial cell line from normal mammary gland, respectively.