Fig 2: ADAM10 mediates platelet aggregation.(A) Representative 2-photon images of C57Bl/6 mice 6–8 hours after P. aeruginosa or S. pneumoniae sepsis, pretreated for 3 days with vehicle or the ADAM10 inhibitor GI254023X. Vasculature (red, Qdots655), platelets (green, GPIbβ). Scale bar: 20 μm. White arrows denote thrombi. (B) Quantification of the total area of platelet accumulation within the vasculature in mouse livers as treated in A. Data represent 5–7 FOV per mouse in 6–7 male and female mice per group and are presented as the mean ± SEM. Measurements for individual FOV within each mouse are displayed in Supplemental Figure 5, A and B. **P ≤ 0.01, by nested t test for each treatment group.

Fig 3: Endothelial ADAM10 mediates lethal sepsis in a pathogen-specific manner.Survival curves for control and VE-Cad ADAM10–/– mice infected with lethal P. aeruginosa (A, n = 21 [13 males, 8 females], n = 20 [11 males, 8 females]); S. pneumoniae (B, n = 24 [8 males, 16 females], n = 23 [11 males, 12 females]; GBS (C, n = 23 [9 males, 14 females], n = 14 [7 males, 7 females]; or C. albicans (D, n = 12 males, 13 males ). **P ≤ 0.01, by log-rank (Mantel-Cox) test for survival curves.

Fig 4: Staphylococcus aureus targets endothelial ADAM10 to cause lethal sepsis.(A) ADAM10 (red) staining in the descending aorta and mesenteric artery from control or VE-Cad ADAM10–/– mice. DAPI (blue) denotes cell nuclei; collagen autofluorescence (green). Scale bars: 30 μm. (B) Survival following lethal S. aureus infection in VE-Cad ADAM10–/– (n = 37) or control (n = 18) female mice. Independent experiments were repeated 4 times, and data were pooled. (C) Survival following lethal purified Hla sepsis in VE-Cad ADAM10–/– (n = 9) or control (n = 10) female mice. Independent experiments were repeated twice, and data were pooled. (D) Serum IL-10 analysis 8 and 24 hours after infection in female VE-Cad ADAM10–/– mice or controls. Data represent 3 independent pooled experiments. Data are presented as the mean ± SEM. (E) Mouse platelet count enumerated 4 hours after lethal infection with S. aureus in male and female mice. Data are from 3 independent pooled experiments and are presented as the mean ± SD. *P ≤ 0.05 and **P ≤ 0.01, by unpaired, 2-tailed t test.

Fig 5: Endothelial ADAM10 mediates platelet aggregation in a pathogen-specific manner.(A) Representative 2-photon images of control and VE-Cad ADAM10–/– mouse livers at 6–8 hours and (B) corresponding quantification of the total area of platelet accumulation within the vasculature of mouse livers after P. aeruginosa, S. pneumoniae, GBS, or C. albicans sepsis. Vasculature (red, Qdots655); platelets (green, GPIbβ). Scale bars: 20 μm. White arrows denote thrombi. Data represent 5–7 FOV per mouse in 5–6 male and female mice per group and indicate the mean ± SEM. Measurements for individual FOV within each mouse are displayed in Supplemental Figure 4, E–H. *P ≤ 0.05 and ***P ≤ 0.001, by nested t test.