Anti cd61 apc

Tetramethylpyrazine (TMP) was obtained from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Ticagrelor (Tica) was from Selleck (Shanghai, China). Adenosine 5'-diphosphate (ADP) and MRS2179 (P2Y₁ receptor antagonist) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Thrombin was from Beijing Solarbio Science & Technology Co., Ltd. (Bejing, China). Dimethyl sulfoxide (DMSO) was from Amresco (Houston, Texas, USA). Antibodies (anti-CD61-APC, anti-CD62P-phycoerythrin-labeled (PE), and anti-GPIIb/IIIa-FITC PAC-1) were purchased from BD Biosciences (San Diego, CA, USA). SQ22536 (an AC inhibitor) and LY294002 (a PI3k inhibitor) were obtained from APExBIO Technology LLC (Houston, Texas, USA). The cyclic AMP enzyme immunoassay (ELISA) kit was from Cayman Chemical (Ann Arbor, MI, USA). The IL-1β and soluble CD40 ligand (sCD40L) ELISA kits were obtained from Elabscience Biotechnology Co., Ltd. (Wuhan, China). The bicinchoninic acid (BCA) protein assay kit was from Beyotime (Shanghai, China). Antibodies against phospho-VASP^ser157, Akt, phospho-Akt^ser473, and phospho-Akt^Thr308 were acquired from Cell Signaling Technology (Beverly, MA, USA). Tetramethylpyrazine, SQ22536, and LY294002 were dissolved in DMSO for further use in experiments. DMSO 0.1% was employed as the vehicle.

Briefly, for the cellular differentiation assay, the cells were seeded at a concentration of 1 × 10⁵ cells/well in 6-well culture plates and incubated with increasing doses of BW18 (5, 10 and 20 µM) for 48 h and 72 h. Cells were washed and resuspended with 100 µL ice-cold PBS. Anti-CD41-FITC, Anti-CD61-APC, Anti-CD71-FITC and Anti-CD235a-APC (BD Biosciences) were, respectively, added to the cell suspension. After 30 min of incubation on ice, cells were washed and resuspended in 500 μL PBS. Quantification of positive cells was analyzed with FACS Calibur flow cytometer (BD Biosciences).