Sterile yellow top tubes

Peripheral blood was collected in acid citrate-dextrose (ACD, 3.2%) sterile yellow-top tubes (Becton, Dickinson and Co.) and was processed within 4 h of collection for all samples. Platelets were isolated by established^{46 (link)–49} purification protocols. Briefly, the ACD-tube whole blood was first centrifuged at 200xg for 20min at room temperature (RT). The platelet-rich plasma (PRP) was removed and Prostaglandin E1 was added to the PRP to prevent exogenous platelet activation. The PRP was then centrifuged at 1000xg for 20min at RT. The platelet pellet was re-suspended in warmed (37 deg C) PIPES saline glucose (PSG). Leukocytes were depleted using CD45⁺ magnetic beads (Miltenyi Biotec). Isolated platelets were further resuspended in Trizol or LDS buffer for RNA (PCR) and protein (Western Blot) analyses.

Peripheral blood was collected in acid citrate-dextrose (3.2%) sterile yellow-top tubes (Becton, Dickinson and Co) and was processed within 4 hours of collection for all samples. Platelets were isolated by established45 (link), 46 (link), 47 (link), 48 (link) purification protocols. Briefly, the acid citrate-dextrose tube whole blood was first centrifuged at 200 g for 20 minutes at room temperature. The platelet-rich plasma (PRP) was removed, and prostaglandin E1 was added to the PRP to prevent exogenous platelet activation. The PRP was then centrifuged at 1000 g for 20 minutes at room temperature. The platelet pellet was resuspended in warmed (37°C) PIPES saline glucose. Leukocytes were depleted using CD45⁺ magnetic beads (Miltenyi Biotec). Isolated platelets were further resuspended in Trizol or LDS buffer for RNA (polymerase chain reaction [PCR]) and protein (western blot) analyses.