ADAP enables kindlin-3 enhancement of αIIbβ3 activation in nonhematopoietic (CHO) cells. CHO cells expressing PAR1, talin, and αIIbβ3 were transiently transfected with mock DNA, ADAP, and/or kindlin-2 (A-B) or kindlin-3 (C-D) along with green fluorescent protein as a transfection marker. Harvested cells were unstimulated or stimulated with 50 μM SFLLRN or 1 mM MnCl2, and specific binding of PAC-1 to green fluorescent protein positive cells, normalized to αIIbβ3 expression, was determined using flow cytometry. (A) Effect of kindlin-2 and ADAP expression on PAC-1 binding (*P < .05). (B) Western blot of lysate from cells in (A) showing expression levels of transfected proteins. (C) Effect of kindlin-3 and ADAP expression on PAC-1 binding (**P < .01). (D) Western blot of lysate from cells in (C) showing expression levels of transfected proteins. Results in (A) and (C) represent the mean ± SEM of 5 experiments.
