LMWH prevents NLRP3 inflammasome activation. (A-B) Inflammasome activation in murine placentas after LMWH and EV injections. Representative immunoblots (A; each lane represents individual mice placentae) and bar graphs (B) showing reduced EV and platelet-induced expression of NLRP3, cleaved IL-1β, and cleaved caspase-1 in murine placenta analyzed at day 12.5 p.c. Control mice, C, were injected with the supernatant obtained after the last PBS wash during EV isolation. (C-E) Inflammasome activation in murine and human trophoblast cells. Representative immunoblots (C) and bar graphs (D, E) showing reduced EV and platelet (PRP) induced expression of NLRP3, cleaved IL-1β, and cleaved caspase-1 due to LMWH in mouse (trophoblast cells [TS] cells C-D) and human (JEG-3, C,E) trophoblasts. Control cells, C, were exposed to supernatant obtained after the last PBS wash during EV isolation. EV represents cells exposed to EV and platelets (PRP); EV+LMWH, cells pretreated with LMWH and exposed to EVs and platelets. (F-H) Representative immunoblots (F) and bar graphs (G-H) showing that LMWH prevented LPS+ATP–induced expression of NLRP3, cleaved IL-1β, and cleaved caspase-1 in mouse (TS cells [F-G]) and human (JEG-3 [F,H]) trophoblasts. Control cells, C, were exposed to PBS. LPS+ATP cells were pretreated with 200 ng/mL LPS (24 hours) and then exposed to 1-mM ATP (1 hour). ATP+LMWH cells pretreated with LMWH (30 minutes), followed by treatment with LPS (24 hour) and then exposed to ATP (1 hour). Inactive (proform) and active (cleaved form) forms of IL-1β or caspase-1 are indicated in panels A,C,F. Only the active form (cleaved form) was quantified in panels B,D-E,G-H. For panels B,D-E,G-H, data shown represent mean ± SEM; n = 5 to 8 placentas from 3 dams for panel B or 3 independent repeat experiments for panels D-E,G-H. ∗P < .05; ns; ANOVA, Šídák multiple comparisons test. EV, EV-injected mice; EV+LMWH, EV-injected mice with LMWH injections.