American Society of Hematology

Figure 6.

$Figure 6. ADAMTS13, but not the mutant without CUB domains, is associated with rafts. MDCK cells stably expressing full-length ADAMTS13-V5-His (A, FL) and the CUB domain–deleted ADAMTS13 mutant (B, delCUB) were lysed with 1% Triton X-100 in MEB buffer, pH 6.5, at 4°C. The cell lysate was subjected to 5% to 40% of sucrose gradient centrifugation as described. Then, 1.2-mL fractions were collected from the top to the bottom. The ADAMTS13 in each faction was determined by Western blotting. Closed arrows indicate intact full-length ADAMTS13 and delCUB mutant. The double asterisks indicate proteolytically degraded ADAMTS13 products. Open arrows indicate caveolin-1 protein (Cav-1), a 21-kDa protein specific for rafts.$

ADAMTS13, but not the mutant without CUB domains, is associated with rafts. MDCK cells stably expressing full-length ADAMTS13-V5-His (A, FL) and the CUB domain–deleted ADAMTS13 mutant (B, delCUB) were lysed with 1% Triton X-100 in MEB buffer, pH 6.5, at 4°C. The cell lysate was subjected to 5% to 40% of sucrose gradient centrifugation as described. Then, 1.2-mL fractions were collected from the top to the bottom. The ADAMTS13 in each faction was determined by Western blotting. Closed arrows indicate intact full-length ADAMTS13 and delCUB mutant. The double asterisks indicate proteolytically degraded ADAMTS13 products. Open arrows indicate caveolin-1 protein (Cav-1), a 21-kDa protein specific for rafts.

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