Fig 2: Specific antibodies inhibited the inflammatory response of the endothelial cells and the expression of LOX-1. (a–d) The expression of LOX-1, MCP-1, and VCAM-1 in the three groups and the corresponding statistical graphs. Values are expressed as mean ± SD. ∗∗P < 0.01 vs. the control group. #P < 0.05 and ##P < 0.01 vs. the Ox-LDL group.

Fig 3: VX765 restrains vascular inflammation and atherosclerosis in low-density lipoprotein receptor–deficient (Ldlr−/−) mice.A Diagramed is the regimen for treatment of Ldlr−/− mice (6–8 weeks old) with VX765 (VX, n = 6) or vehicle (DM, n = 6). B, C The atherosclerotic lesions in the entire aortas (n = 6) and the aortic sinuses were visualized by oil red O staining. D The necrotic areas were visualized by Verhöeff staining. E–J The expression of the indicated proteins in the aortic sinuses was detected by immunohistochemical staining. The positive area of each protein was measured. K–N Dual immunofluorescence staining of the indicated proteins in the aortic sinuses. O–Q The expression of Caspase 3 and Mac3 was stained (O) and quantitated to assess cell death (P) and in situ macrophage efferocytosis (Q). R Profile of lipids and lipoproteins (n ≥ 3). HFD high-fat diet; ip intraperitoneal; LDL low-density lipoprotein; HDL high density lipoprotein; TC total cholesterol; TG total triglyceride; DM DMSO; VX VX765; LC3 microtubule-associated protein light chain 3; α-SMA α-smooth muscle actin; MCP-1 monocyte chemoattractant protein-1; ICAM-1 intercellular cell adhesion molecule 1; Casp1 cleaved caspase 1; Casp3 cleaved caspase 3; IL-1β interleukin 1β; ACs apoptotic cells; ns no significance; *p < 0.05; **p < 0.01.

Fig 4: NLRP3 deficiency impairs the ability of VX765 to manipulate diet-induced vascular inflammation and atherosclerosis in ApoE−/− mice.A Diagramed is the regimen for treatment of ApoE−/− mice with VX765 (VX, n = 4) or vehicle (DM, n = 5) and Nlrp3−/−;ApoE−/− mice with VX765 (VX, n = 7) or vehicle (DM, n = 9). B The atherosclerotic lesions in the whole aortas were visualized by oil red O staining (n ≥ 3). C The necrotic regions in the aortic sinuses were visualized by Verhöeff staining (n ≥ 3). D–I The expression of the indicated proteins in the aortic sinuses was determined by immunohistochemical staining. The positive area of each protein was measured (n ≥ 3). J–O Immunostaining analysis of the aortic sinuses with the indicated antibodies. Lesional macrophage expression of LC3 and p62 were quantitated. P–R The expression of caspase 3 and Mac3 was stained and quantitated to measure cell death and in situ macrophage efferocytosis. S Measurement of IL-1β content in the blood from ApoE−/− and Nlrp3−/−ApoE−/− mice treated as indicated in (A) by ELISA (n = 3). T Schematic model for the role and mechanisms of VX765 action. DKO Nlrp3−/−;ApoE−/− mice; HFD high-fat diet; ip intraperitoneal; DM DMSO; VX VX765; α-SMA α-smooth muscle actin; MCP-1 monocyte chemoattractant protein-1; ICAM-1 intercellular cell adhesion molecule 1; LC3 microtubule-associated protein light chain 3; IL-1β interleukin 1β; MΦ macrophage; ACs apoptotic cells; ns no significance; *p < 0.05; **p < 0.01.

Fig 5: VX765 restricts vascular inflammation and atherosclerosis in apolipoprotein E–deficient (ApoE−/−) mice.A Diagramed is the regimen for treatment of ApoE−/− mice (6–8 weeks old) with VX765 (VX, n = 11) or vehicle (DM, n = 11); B, C The atherosclerotic lesions in the whole aortas (n = 11) and the aortic roots (n = 4) were visualized by oil red O staining. D The necrotic regions were visualized by Verhöeff staining. E–J The expression of the indicated proteins in the aortic sinuses was detected by immunohistochemical staining. The positive area of each protein was measured. K The profile of lipids and lipoproteins (n = 3). L, M Double immunostaining of the indicated proteins in the aortic sinuses. N–S The expression of Caspase 3 and Mac3 was stained and quantitated to assess cell death and in situ macrophage efferocytosis. T–W Macrophage autophagy was assessed by immunostaining with the indicated antibodies and quantitated. HFD high-fat diet; ip intraperitoneal; LDL low-density lipoprotein; HDL high density lipoprotein; TC total cholesterol; TG total triglyceride; α-SMA α-smooth muscle actin; MCP-1 monocyte chemoattractant protein-1; ICAM-1 intercellular cell adhesion molecule 1; IL-1β interleukin 1β; Casp1 cleaved caspase 1; Casp3 cleaved caspase 3; ACs apoptotic cells; DM DMSO; VX VX765; MΦ macrophages; LC3 microtubule-associated protein light chain 3; ns no significance; *p < 0.05; **p < 0.01; ***p < 0.001.