Fig 1: rTFPI and thiol blockers prevent S-proteins-induced thrombin expression and release in HLMEC, whereas TM had no effect: (A,B) HLMEC treated (24 h) with S-proteins (1 nM SW or SD), with or without rTFPI, DTNB, and BDCA3 (200 ng/mL); (C) HLMEC were treated 6 h with S-proteins, with or without rTFPI, DTNB, and BDCA3, washed, and co-cultured with neutrophils for 24 h; (D) human neutrophils were treated 6 h with S-proteins, with or without rTFPI, DTNB, and BDCA3, washed, and co-cultured with HLMEC for 24 h. Following treatments, thrombin levels in culture supernatants (A,C,D) and endothelial cells lysates (B) were quantified by ELISA. Data presented as mean ± standard deviation. Control: untreated cells; Hi: heat-inactivated (SW, SD, rTFPI, DTNB, BDCA3). # p < 0.0001.
Fig 2: rTFPI and thiol blockers prevent S-protein-induced Factor-V expression and release in HLMEC, whereas TM had no effect: (A,B) HLMEC treated (24 h) with S-proteins (1 nM SW or SD), with or without rTFPI, DTNB, and BDCA3 (200 ng/mL); (C) HLMEC were treated 6 h with S-proteins, with or without rTFPI, DTNB, and BDCA3, washed, and co-cultured with neutrophils for 24 h; (D) human neutrophils were treated 6 h with S-proteins, with or without rTFPI, DTNB, and BDCA3, washed, and co-cultured with HLMEC for 24 h. Following treatments, Factor-V levels in culture supernatants (A,C,D) and endothelial cells lysates (B) were quantified by ELISA. Data presented as mean ± standard deviation. Control: untreated cells; Hi: heat-inactivated (SW, SD, rTFPI, DTNB, BDCA3). # p < 0.0001.
Fig 3: rTFPI, TM, and thiol blockers prevent S-proteins-induced fibrinogen expression and release in HLMEC: (A,B) HLMEC treated (24 h) with S-proteins (1 nM SW or SD), with or without rTFPI, DTNB, and BDCA3 (200 ng/mL); (C) HLMEC were treated 6 h with S-proteins, with or without rTFPI, DTNB, and BDCA3, washed, and co-cultured with neutrophils for 24 h; (D) human neutrophils were treated 6 h with S-proteins, with or without rTFPI, DTNB, and BDCA3, washed, and co-cultured with HLMEC for 24 h. Following treatments, fibrinogen levels in culture supernatants (A,C,D) and endothelial cell lysates (B) were quantified by ELISA. Data presented as mean ± standard deviation. Control: untreated cells; Hi: heat-inactivated (SW, SD, rTFPI, DTNB, BDCA3). # p < 0.0001.
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