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. 2021 Dec 6;10(12):3425. doi: 10.3390/cells10123425

Figure 3.

Figure 3

Giardia-induced ROS/NLRP3/CASP1/GSDMD-mediated macrophage pyroptosis. PMs were pretreated with 10 μM NAC, 40 μM AYC, or 50 μM MCC950 for 2 h, and stimulated with trophozoites at a ratio of 10 parasites/cell for 12 h. (A,B) The effects of NAC or AYC pretreatment on Giardia-induced upregulation of NLRP3, cleavage of CASP1 and GSDMD, and secretion of IL-1β and IL-18 were analyzed by Western blotting. (C,D) The effects of MCC950 pretreatment on Giardia-induced cleavage of CASP1 and GSDMD and secretion of IL-1β and IL-18 were analyzed by Western blotting. (E) After pretreatment with NAC, AYC, or MCC950, ASC expression in Giardia-treated PMs was detected by immunofluorescence assay (scale bar = 100 μm). (F,G) NAC, AYC, or MCC950 pretreatment affected the Giardia-induced decrease in cell viability and increase in LDH release. All experiments were repeated at least three times. (A,C,E) Representative pictures are shown. (B,D,F,G) Values are expressed as the mean ± SD (** p < 0.01 as compared to nontreated controls or as indicated). GiTr: reads Giardia trophozoites, and ns: no significant difference.