Western blot analysis of ALCAM expression in lysate and conditioned medium of TPC-1 cell line.

<p>(A) Analysis of TPC-1 cell lysate (Lys) and conditioned medium (CM) using anti-ALCAM antibody. Arrow indicates membrane-localized ALCAM isoform in lysate, and arrowheads indicate shed-ALCAM isoforms in CM, respectively. The SKOV-3 cell line was used as positive control. (B) TPC-1 cell lysate and CM were treated or not with a mixture of O-glycosidase (O) and sialidase (S) and analyzed by western blot using anti-ALCAM antibody. Continuous arrows indicate membrane and shed ALCAM forms before digestion, and dashed arrows indicate digestion products. (*) indicates less evident bands of ALCAM in CM. (C) TPC-1 cell lysate and CM were treated or not with O-glycosidase (O), sialidase (S), or PNGaseF (F) and analyzed by western blot using anti-ALCAM antibodies. Continuous arrows indicate membrane and shed ALCAM forms before digestion, and dashed arrows indicate digestion products.