Antithrombotic effect of BZ is KLF2 dependent. (A) Endothelial (human umbilical vein endothelial cells [HUVECs]), monocytic (RAW 264.7), and megakaryocyte (MEG-01) cell lines treated with BZ at 5 µM for 24 hours (*P < .05; n = 3 to 5 mice per group). Total RNA was isolated, and KLF2 mRNA expression was analyzed by quantitative polymerase chain reaction (qPCR) and was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). (B) C57BL/6J mice were treated with BZ, and WBCs were isolated for KLF2 mRNA expression and were normalized to GAPDH (*P < .05; n = 5). (C) Carotid artery thrombosis assay (photochemical injury model) was performed in CAG-Cre-ERT²  (CRE), GL-K2-KO, and GL-K2-TG mice. (D) CRE and GL-K2-KO mice were treated with intraperitoneal BZ (0.3 mg/kg) vs saline 3 times per week for 2 weeks followed by carotid artery thrombosis assay (photochemical injury model) performed 24 to 30 hours after the last dose of BZ. (E) Total RNA was isolated from primary KLF2 knockout and overexpressed macrophages (peritoneal macrophages) and endothelial cells (cardiac and pulmonary), and mRNA expression was analyzed as indicated by qPCR and normalized to GAPDH. eNOS, endothelial nitric oxide synthase; TF, tissue factor; WT, wild-type.