Anemia / 2012 / Article / Fig 1

Research Article

Sickle Cell Disease Activates Peripheral Blood Mononuclear Cells to Induce Cathepsins K and V Activity in Endothelial Cells

Figure 1

Sickle cell disease preconditions circulating peripheral blood mononuclear cells to induce cathepsin K activity. Whole blood samples were obtained from donors homozygous for the normal β-globin allele (AA) and homozygous for the sickle allele (SS). (a) Baseline serum levels of TNFα were quantified using an ELISA specific for human TNFα ( , , SEM bars shown). (b) PBMCs were isolated via differential centrifugation through a density gradient. For cocultures, confluent EC cultures were preconditioned with 10 ng/mL TNFα for 4 hours, prior to the addition of either AA or SS PBMCs. Nonadherent cells were washed away, and cocultures were maintained for an additional 20 hours. Representative images of cocultures were used for mononuclear cell adhesion counts. (c) Cells were lysed and cathepsin K activity was assessed using multiplex cathepsin zymography and quantified via densitometry ( , ).

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