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Fig. 1. Identification of rat {alpha}6ß4. (A) Lysate of Progressor cells was purified by immunoaffinity chromatography and separated by SDS-PAGE, which revealed two bands of 130 and 200 kDa. The trypsin digest of these 2 bands was subjected to mass spectrometry. (B) PCR analysis of the metastasizing lines BSp73ASML, Progressor and Regressor and of the non-metastasizing line BSp73AS with primers (see Materials and Methods) detecting the 7 AS deletion (ß4D) (left) and the +53 and/or +70 insertion (ß4B/ß4C) (right) of the ß4 integrin. The 3 metastasizing lines express the ß4A isoform. The non-metastasizing line does not express the ß4 integrin. (C) Staining of BSp73ASML, BSp73AS and BSp73AS-ß4 cells confirms that B5.5 recognizes {alpha}6ß4. Black line: negative control (secondary antibody), light gray area: staining with B5.5, dark gray area: staining with anti-ß1. Single parameter overlays are shown.