Fig 2: Thrombosis is delayed in CDH6 knockout mice. Thrombosis was initiated in Cdh6 −/− and wild‐type mice using the (A) FeCl3 (n = 9 per group) or (B) Rose Bengal (n = 5 for wild‐type and n = 6 for Cdh6−/−) models and time to vessel occlusion was observed. The vessels were monitored until a full occlusion was formed or the experiment ended and represented as Kaplan‐Meier curves. The curves were compared using the Mantel‐Cox log‐rank test. (C) Platelet count was measured in whole blood from wild‐type and Cdh6 −/− mice and compared using Student's t test. (D) Cdh6 −/− and wild‐type mice were anesthetized before clipping 3 mm of tail to measure time to bleeding cessation; significance was evaluated with Student's t test, and bars represent mean ± SEM

Fig 3: Human platelets express cadherin-6. (A) Human platelet lysate was probed with mouse monoclonal anti–cadherin-6; CHO cells transfected with human cadherin-6 (Cdh6) was used as a control. A standard curve was generated using recombinant human cadherin-6 Fc chimera protein. The blot shown is representative of n = 2. (B) Human platelet lysate was immunoprecipitated with anti-cadherin-6 antibody. Cadherin 6, a-catenin, and ß-catenin were detected by western blot. (C) Quantitative flow cytometry was performed using an APC-conjugated anti-cadherin-6 antibody on nonstimulated platelets or those activated with PAR1 (SFLLRN, 25 µmol/L) or PAR4 (AYPGKF, 250 µmol/L) stimulating peptides, ADP (5 µmol/L), or convulxin (5 nmol/L). CHO, Chinese hamster ovary; PAR, protease-activated receptor; PLTs, platelets

Fig 4: Blocking cadherin-6 disrupts thrombosis. C57BL/6J mice were treated with sheep polyclonal anti–cadherin-6 (Cdh6, 0.8 mg/kg) 15 minutes before initiating thrombosis with FeCl3 or Rose Bengal. (A) Intravital microscope images obtained following FeCl3 treatment. In IgG-treated mice at 8 minutes, the proximal end of the thrombus was monitored to accurately detect full occlusion. Time to stable vessel occlusion was observed and represented as Kaplan-Meier curves for (B) FeCl3 (n = 3 per group) and (C) Rose Bengal models (n = 5 for no treatment, n = 4 for IgG or Cdh6). The curves were compared using the Mantel-Cox log-rank test