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. 2000 Apr 17;191(8):1413–1422. doi: 10.1084/jem.191.8.1413

Figure 2.

Figure 2

Affinity isolation of PSGL-1 glycoprotein from platelets and neutrophils. (A) Purified preparations of neutrophils or platelets were biotinylated and lysed. Cell lysates were incubated with P-selectin–IgG (P-sel-IgG) chimera or anti–PSGL-1 (PSL-275). Specifically bound proteins were eluted and separated on 7.5% SDS gel under nonreduced conditions. (B) PSGL-1 was immunopurifed from neutrophil and platelet lysates. SDS-eluted proteins were analyzed on 7.5% SDS gel. The blotted proteins were detected with P-selectin–IgG and HRP-conjugated anti–human IgG. (C) Biotinylated platelets were lysed and subjected to affinity isolation with P-selectin–IgG (P-sel-IgG). The high molecular weight complex (*) seen in the nonreduced (NR) sample likely represents a disulfide-bonded multimer formed during platelet lysis through exposure of free sulfhydryl from the transmembrane and the cytoplasmic domains of PSGL-1. Note that the major species observed in the reduced lane (R) is of the expected size 120 kD for PSGL-1 monomer (arrow). The numbers on the side indicate migration of marker proteins in kD.