Fig. 4
NEMO is required for the regulation of NEK7/NLRP3 function, oxidative stress and iron overload in Caspase 6-deficient livers in response to IR stress. A Schematic figure for the injection of mannose-conjugated NEMO siRNA delivery system into ischemic livers of Caspase 6KO mice. B Immunofluorescence staining of AlexaFluor488-labeled CD68 positive macrophages and AlexaFluor Cy5-labeled control siRNA in ischemic livers, Scale bars, 20 μm, 10 μm, N = 4/group. C Liver NEMO expression was evaluated by Western blot assay, N = 6/group. D Serum ALT/AST level in ischemic livers, N = 6/group. E H&E staining of ischemic livers and Suzuki’s scores. Scale bar: 100 μm, N = 6/group. F Immunofluorescence staining and quantification of CD11b+ macrophages in ischemic livers, Scale bar: 50 μm, N = 4/group. G IHC staining and quantification of Ly6G.+ neutrophils in ischemic livers, Scale bar: 50 μm, N = 4/group. H Western blotting analysis and relative intensity of NEK7 and NLRP3, N = 6/group. I Detection of cytokines IL-1β, TNF-α and CXCL-10 by qRT-PCR in ischemic livers, N = 6/group. ELISA analysis of serum IL-1β levels (J) and serum HMGB1 levels (K), N = 6/group. L Hepatic MDA concentration in ischemic livers, N = 6/group. M 4-HNE staining in IR-induced liver tissues, scale bar: 50 μm, N = 4/group. N Non-heme iron profiles in liver tissues by colorimetric assay, N = 6/group. All data represent the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001