GST-talin and GST-kindlin-1 pull-downs of ADAP and its truncation mutants. (A) Schematic map of ADAP truncation mutants. (B-E) CHO cells expressing PAR1, talin, and αIIbβ3 were transfected with the indicated FLAG-tagged ADAP (FLAG-ADAP) mutant constructs and harvested after 48 hours, and lysates were subjected to pull-down assays. (B) GST-kindlin-1 and GST pull-downs were probed for FLAG (ADAP). Gels were stained with Coomassie to assess loading, and blots were reprobed for β3 as a positive control. (C) Western blot of lysates from transfected cells in (B) showing truncated FLAG-ADAP protein expression. (D) Western blot of GST-talin and GST pull-downs probed for FLAG (ADAP). (E) Western blot of lysates from transfected cells in (D) showing FLAG-ADAP protein expression. Results are representative of at least 4 experiments. See the text for interpretations.