Figure 3.

Different effects of cell release by trypsin and NECDS on subsequent initiation of anoikis and on antibody accessibility to the cell surface anoikis-initiating molecules in MUC1-positive and -negative cells. (a) HCA1.7+/− cells were released by NECDS or trypsin and cultured in suspension for 24 h before the cell-associated caspase-3/-7 activities were assessed. The data are presented as mean±S.E.M. of triplicate determinations of two independent experiments. (b–d) Representative flow cytometry plots show antibody access to cell surface MUC1 (b), E-cadherin, Integrinβ1, CD44 (c, d) and recombinant Fas-L access to cell surface Fas (f) in HCA1.7+/− cells released by trypsin or NECDS. Note, an additional integrinβ1 peak (arrowed) is seen in HCA1.7− cells released by NECDS in comparison to those released by trypsin. (e) Immunoblotting of cell lysates shows total cellular expression of CD44, E-cadherin, integrinβ1, Fas, Fas-L and tubulin in HCA1.7+/− cells