Figure 2.

Identification and characterization of actin as a triggering receptor expressed on myeloid cells-1 (TREM-1)-interacting protein on platelets. (A) SDS-PAGE gel analysis of the proteins extracted from platelets which were co-purified with recombinant extracellular domain of mouse TREM-1 (rTREM-1) (fusion with FLAG and 6His tag) through nitrilotriacetic acid (NTA) agarose resin and αFLAG-agarose. The interacting protein band was marked by an arrow. (B) LC-MS/MS analysis of the protein band extracted from the SDS-PAGE gel (A). The amino acid marked by yellow was identified by MS/MS characterization. (C) The purified proteins were further characterized by immunoblotting with antibody against actin or TREM-1. (D) The rACTIN purified with NTA agarose resin was incubated with rTREM-1 (fusion with FLAG and 6His tag) purified with NTA agarose resin and then loaded on the αFLAG-agarose. The elution was analyzed by immunoblot with antibody against actin or TREM-1.