FIGURE 4.

Screen of MDCK-expressed MUC1 binding to recombinant canine galectins. MDCK cells stably expressing MUC1 were pulse labeled for 30 min with [³⁵S]Met/Cys and chased for 90 min prior to immunoprecipitation of MUC1 from cell extracts. Resuspended immunoprecipitates were incubated with individual GST-galectins and glutathione-conjugated beads and eluted with sucrose (S, control), then lactose (L, specific binding) and then glutathione (for Coomassie stain of GST-Gal) and subjected to SDS-PAGE and transferred to nitrocellulose. A representative profile of [³⁵S]MUC1 (A), immunoblotted large or small subunits of MUC1 (B), and glutathione-eluted GST-Gal stained with Coomassie (C) are shown. Analysis of multiple experiments is shown in Fig. 5.