Fig. 3: Vagus nerve stimulation requires α7nAChR to increase platelet cytosolic calcium, enhance cellular activation, and accelerate local clot formation to reduce bleeding.

a C57BL6/J mice received vagus nerve stimulation or sham stimulation before harvesting circulating platelets to measure basal cytosolic calcium uptake. Data were presented as mean ± s.e.m. Sham (n = 5) and VNS (n = 6) mice per group. **p = 0.0073 vs. sham. Statistical significance was determined by unpaired two-tailed Student’s t-test. The figure represents pooled results from two experiments performed independently. b α7nAChR-deficient (α7KO) mice received vagus nerve stimulation or sham stimulation before harvesting circulating platelets to measure basal cytosolic calcium uptake. Data were presented as mean ± s.e.m. Sham (n = 4) and VNS (n = 4) mice per group. p = 0.32 vs. sham. Statistical significance was determined by unpaired two-tailed Student’s t-test. The figure represents pooled results from two experiments performed independently. c C57BL6/J mice received vagus nerve stimulation or sham stimulation before harvesting platelets for analysis of P-selectin (CD62P) expression. Data were presented as mean ± s.e.m. Sham (n = 13) and VNS (n = 12) mice per group. p = 0.98 vs. sham. Statistical significance was determined by unpaired two-tailed Student’s t-test. The figure represents pooled results from three experiments performed independently. d Representative FACS histogram of platelet P-selectin (CD62P) expression after vagus nerve stimulation or sham stimulation. Experiments were performed independently three times with similar results. e C57BL6/J mice received vagus nerve stimulation or sham stimulation before harvesting platelets, ex vivo thrombin stimulation, and analysis of P-selectin (CD62P) expression. Data were presented as mean ± s.e.m. Sham (n = 10) and VNS (n = 12) mice per group. *p = 0.029 vs. sham. Statistical significance was determined by unpaired two-tailed Student’s t-test. The figure represents pooled results from three experiments performed independently. f Representative FACS histogram of platelet P-selectin (CD62P) expression after vagus nerve stimulation or sham stimulation followed by ex vivo thrombin stimulation. Experiments were performed independently three times with similar results. g α7nAChR-deficient (α7KO) mice received vagus nerve stimulation or sham stimulation before harvesting platelets, ex vivo thrombin stimulation, and analysis of P-selectin (CD62P) expression. Data were presented as mean ± s.e.m. Sham (n = 11) and VNS (n = 11) mice per group. p = 0.57 vs. sham. Statistical significance was determined by unpaired two-tailed Student’s t-test. The figure represents pooled results from three experiments performed independently. h Platelets from uninjured C57BL6/J mice receiving vagus nerve stimulation or sham stimulation were transferred into naïve animals before tail transection. Data were presented as mean ± s.e.m. Sham (n = 9) and VNS (n = 8) mice per group. *p = 0.016 vs. sham. Statistical significance was determined by unpaired two-tailed Student’s t-test. The figure represents pooled results from two experiments performed independently. i C57BL6/J mice received vagus nerve stimulation or sham stimulation before tail transection and collection of systemic or local shed blood for measurement of (TAT) complexes. Data were presented as mean ± s.e.m. Sham VNS + Systemic (n-4), VNS + Systemic (n = 4), Sham VNS + Local (n = 3), VNS + Local (n = 3) mice per group. **p = 0.003 vs. sham in local blood. p = 0.42 VNS vs. sham in systemic blood. Statistical significance was determined by unpaired two-tailed Student’s t-test. The figure represents pooled results from two experiments performed independently. j Representative low power (left, scale bar = 100 µm) and high power (right, scale bar = 50 µm) lung images from C57BL6/J mice after sham stimulation and tail transection. Experiments were performed independently three times with similar results. k Representative low power (left, scale bar = 100 µm) and high power (right, scale bar = 50 µm) lung images from C57BL6/J mice after vagus nerve stimulation and tail transection. Experiments were performed independently three times with similar results. Source data are provided as a Source Data File.