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. 2022 Sep 5;12:989167. doi: 10.3389/fonc.2022.989167

Figure 6.

Figure 6

IVM directly interact with GDSMD to promote its oligomerization, and subsequent GSDMD-dependent NETs. (A) Western blot analysis of Caspase-11, GSDMD and cit-H3 expression in bone marrow neutrophils from control, B16F10 and B16F10 with IVM group. (B) Quantitative analysis of caspase-11, GSDMD-N and cit-H3 in (A, C) Non-reducing western blot analysis of GSDMD in bone marrow neutrophils after CTSB or CTSB with IVM treatment. (D) Quantitative analysis of GSDMD oligomer. (E) Reducing western blot analysis of GSDMD in bone marrow neutrophils isolated from WT mice treated with CTSB for 12 h followed by treatment with IVM for 2 h. (F) Quantitative analysis of GSDMD-N. (G) and (H) MST analysis determined the K d of IVM towards His-GSDMD (267.96 nm) (G) or His-Caspase-4 (poor fitting) (H) labeled with RED-tris-NTA 2nd Generation dye. Concentration is reported in nanomolar. Data in all panels are representative of at least three independent experiments. *P < 0.05; ns, no significance [two-way ANOVA in (B), one-way ANOVA in (F)].