American Society of Hematology

Fig. 3.

$Fig. 3. Detection of CD63 and GPIb in platelet lysate, microvesicles, and exosomes. (A) Platelet releasates were centrifuged at 750g (PLlys) and 10,000g(104-g) after stimulation with 15 μmol/L TRAP. The exosome-enriched fraction was obtained after centrifugation of the supernatant at 65,000g. The membrane pellet was dissolved and then floated into a linear sucrose gradient. Top-bottom gradient fractions were diluted in SDS-sample buffer and analyzed by Western blotting for the presence of GPIb and CD63. (B) Relative distribution of CD63 and GPIb quantified from (A). CD63 is enriched in the fractions 5 and 6 from the sucrose gradient, equilibrating at density ∼1.16 g/mL. The majority of GPIb is recovered in PLlys, and the 104-g fraction and somewhat overlapping in the exosome fraction. PLlys, platelet pellet; 104-g, microvesicle fraction.$

Detection of CD63 and GPIb in platelet lysate, microvesicles, and exosomes. (A) Platelet releasates were centrifuged at 750g (PL_lys) and 10,000g(10⁴-g) after stimulation with 15 μmol/L TRAP. The exosome-enriched fraction was obtained after centrifugation of the supernatant at 65,000g. The membrane pellet was dissolved and then floated into a linear sucrose gradient. Top-bottom gradient fractions were diluted in SDS-sample buffer and analyzed by Western blotting for the presence of GPIb and CD63. (B) Relative distribution of CD63 and GPIb quantified from (A). CD63 is enriched in the fractions 5 and 6 from the sucrose gradient, equilibrating at density ∼1.16 g/mL. The majority of GPIb is recovered in PL_lys, and the 10⁴-g fraction and somewhat overlapping in the exosome fraction. PL_lys, platelet pellet; 10⁴-g, microvesicle fraction.

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