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. Author manuscript; available in PMC: 2020 Jun 7.
Published in final edited form as: Circ Res. 2019 Apr 22;124(12):1747–1759. doi: 10.1161/CIRCRESAHA.119.314845

Figure 4. MAC-induced NLRP3 inflammasome induces IL-1β maturation and gasdermin-D-dependent IL-1β release from human ECs.

Figure 4.

(A) IFN-γ-primed ECs were pre-treated with NLRP3 inhibitor MCC950 prior to PRA sera treatment and immunoblot analysis of whole cell lysates for cleaved caspase-1 and gasdermin D amino terminal region (GSDMD-N) and culture supernatants for cleaved IL-1β. (B) IFN-γ-primed ECs were pretreated with selective caspase-1 inhibitor z-YVAD-FMK prior to PRA sera treatment and immunoblot analysis of whole cell lysates for cleaved caspase-1 and GSDMD-N and culture supernatants for cleaved IL-1β. (C) ELISA measurements of IL-1β in culture supernatants of IFN-γ-primed ECs pretreated with z-YVAD-FMK or DMSO prior to PRA treatment and compared to IFN-γ-primed ECs simply washed with PRA sera (t=0 control) (n=3, Student t-test, SEM). (D) IFN-γ-primed ECs were transfected with either control or gasdermin D siRNA and whole cell lysates were assessed for cleaved caspase-1 and culture supernatants were assessed for cleaved IL-1β. **P<0.01.