Figure 7.
The C2-24 disulfide bond is required for β3 homophilic binding in transfected cells. A, B) HEK293 cells were cotransfected with plasmids expressing β3-wt-EGFP and β3-Ig-6His (A), or cotransfected with β3-[C24A]-EGFP and β3-Ig-6His (B). Samples were precipitated with Talon beads, and blotted as indicated. C) HEK293 cells were separately transfected with plasmids expressing β3-wt-EGFP and β3-wt-Myc. Cells were cocultured for 2 d under conditions of 70–90% confluency prior to cell lysis. Samples were immunoprecipitated with anti-Myc agarose and blotted as indicated. D) Cells were separately transfected with plasmids expressing β3-[C24A]-EGFP and β3-wt-Myc, then cells were cocultured for 2 d under conditions of 70–90% confluency prior to cell lysis and immunoprecipitation with anti-Myc agarose and blotting as indicated. FT, flow through; W, wash; E, elution fraction.