Fig. 8.

Deletion of α_IIb CT MD region reduced α_IIb and β₃ transmembrane domain association but did not affect the binding of the talin-1 head. (A) Disulfide crosslinking of indicated α_IIbβ₃ constructs expressed in 293FT cells. Immunoprecipitates of [³⁵S]-labeled integrins were subjected to non-reducing SDS-PAGE and autoradiography (upper panel). The quantification of crosslinking efficiency is shown in the lower panel. Data are presented as mean±s.e.m (n≥3). (B) PAC-1 binding of 293FT cells transfected with indicated α_IIbβ₃ constructs in the buffer containing 1♣mM Ca²⁺ and 1♣mM Mg²⁺ (Ca/Mg) or 0.2♣mM Ca²⁺ plus 2♣mM Mn²⁺ (Ca/Mn). (C) Co-immunoprecipitation of the talin-1 head with α_IIbβ₃ integrin. 293FT cells were co-transfected with α_IIbβ₃ constructs and EGFP or EGFP–TH. Integrins were immunoprecipitated (IP) with mAb 10E5 and subjected to western blotting (WB) with anti-EGFP and anti-β₃ antibodies.