Fig. 6. Transfection of full-length Bves promotes aggregation in L-cells. L-cells were transfected with full-length Bves (B, L-F cells) or full-length Bves with an engineered Kozak sequence (C, L-K cells). Aggregation assays were performed on transfected and non-transfected cells as described in Materials and Methods and the degree of adhesion was calculated. Non-transfected cells did not aggregate (A). Graphs show that both L-F cells and L-K cells gained adhesive activity significantly over controls. (D-E) Bves-expressing cells are distinct from the non-transfected cells. Equal numbers of non-transfected L-cells were labeled with DiI, and Bves-transfected L-K cells were mixed and allowed to aggregate. Cells were stained with DAPI to visualize all cells. In the three examples given, note that aggregates are composed almost exclusively of transfected cells. (G,H) Promotion of aggregation in L-K cells was reduced in the presence of blocking antibody. When blocking antibody was added at 1:100 dilution to the cell suspension, reduction of aggregates were observed (see graph). Increase of antibody concentration (1:50 dilution) results in decrease in aggregates (H, graph) compared with aggregates without blocking antibody (G), suggesting that aggregation of L-cells results in expression of Bves. The ratio of aggregate to total cell is given with the s.e.m.