Fig. 6.
Inhibition of NLRP3 deubiquitylation decreases inflammasome activation and atherosclerosis. Ldlr−/− mice were transplanted with BM from Myl-Abcdko mice and fed WTD. After 2 weeks WTD feeding, mice were injected intraperitoneally with HL (1 mg/kg) or vehicle control (DMSO/PBS) for 6 weeks. Mice were sacrificed after a total of 8 weeks of WTD. A, B: Immunoblot of intracellular Caspase-1 and GSDMD cleavage in Ly6G−CD11b+splenic monocytes/macrophages (A) and splenic Ly6G+ neutrophils (B). C: Atherosclerotic lesion area and necrotic core area in the aortic root. Representative images of hematoxylin and eosin (H&E)–stained sections are shown; atherosclerotic plaques are delineated by dashed lines. Scale bars, 200 μm. D: Fibrous cap thickness analysis via a Masson trichrome staining. Bracketed regions show fibrous cap thickness of aortic root plaques. Scale bars, 100 μm. E, F: Neutrophils were stained in atherosclerotic lesions using Ly6G (E) or MPO (F), and Ly6G+ (E) or MPO+ (F) percentages of lesion size was quantified. Concomitantly, lesions were stained for 3HCit. To assess NETs, the overlap of Ly6G and 3HCit (E) or MPO and 3HCit (F) was quantified as percentage of the total lesion area. Each datapoint represents an individual mouse. Scale bars, 50 μm. N = 20. ∗P < 0.05, by t test.