Fig 1: 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 2: Quantitative real-time PCR and Western blot analysis of SR-BI, ABCG5, and ABCG8. A Quantitative real-time PCR analysis of the mRNA levels of SR-BI, ABCG8 and ABCG5 in liver of the mice in the control and LOX-1 groups. The expression of mRNA was normalized to that of GAPDH. B-E Western blot analysis of the hepatic lipid metabolism core pathway-related proteins SR-BI, ABCG5, and ABCG8 8 weeks after virus transduction. Values are expressed as mean ± SD. **P < 0.01 compared with the control group
Fig 3: Model summarizing the proposed effects of differences in mTRIB1 expression on foam cell expansion in early-stage atherosclerosis.Factors up-regulating mTRIB1 expression in human MDMs and plaque macrophages simultaneously increase cholesterol and neutral lipid uptake, with no compensatory rise in cholesterol efflux. Schematic recognizes that increased OLR1 expression increases the probability of this scavenger receptor assembling as a hexamer made up of three homodimers on the macrophage cell surface and that this configuration leads to a marked increase in its affinity for oxLDL (9) and hence OLR1-mediated uptake of oxLDL lipids. In the setting of no compensatory rise in HDL-mediated cholesterol efflux, accelerated foam cell expansion and increased atheroma burden ensue, highlighting the therapeutic potential of inhibiting macrophage Trib1 expression to block the gene expression changes that both promote macrophage cholesterol uptake and cholesteryl ester formation accumulation and prevent the increased hydrolysis of the accumulated cholesteryl ester and thus the up-regulation of the reverse cholesterol transport pathway to mediate the removal of cholesterol from the arterial wall.
Fig 4: The immune intervention with the Ox-ApoB peptide fragment protects the vascular endothelial barrier connection, reduces the expression of LOX-1 and cell apoptosis in the plaque, and increases the clearance of Ox-LDL by liver macrophages. (a, d) Representative images of aortic root sections obtained from the control and immunization groups stained with ZO-1 and CD31 (a). Nuclei were stained with DAPI (blue). Compared with the control group (set as 1), the relative ratio of CD31 that was occupied by ZO-1 is presented among the indicated groups (d). (b, e, f) Representative images of aortic root sections obtained from the control and immunization groups stained with TUNEL and LOX-1 (b). Nuclei were stained with DAPI (blue). The percentage of the plaque area that was occupied by TUNEL and LOX-1 cells is presented among the indicated groups (e, f). (c, g) Representative images of liver sections obtained from the control and immunization groups stained with ABCA1 and F4/80 (c). Compared with the control group (set as 1), the relative ratio of F4/80 occupied by ABCA1 is presented among the indicated groups (g). Data are expressed as the mean ± SD. ∗P < 0.05, ∗∗P < 0.01.
Fig 5: Myeloid TRIB1 induces reciprocal changes in oxLDL and HDL receptor expression in human and mouse macrophages.(A) TRIB1 RNA levels in monocytes and MDMs of CTS participants (24). Correlation (R2 < 0.001, P = 0.47) was performed on 596 paired monocyte and MDM samples. (B) MDM (n = 596) and monocytes (n = 758) were ranked according to TRIB1 RNA contents and gene expression values in the top and bottom quartiles compared. Log2 fold changes (FC) of specified RNAs in TRIB1High (n = 149) versus TRIB1Low (n = 149) MDM, with associated P values. (C) FC and P values for differential expression of RNAs encoding representative scavenger receptors, including CD36, which mediates (ox)-phospholipid and long-chain fatty acid uptake (39), the acetylated-LDL scavenging receptor (40), and macrophage scavenger receptor (40). Comparisons are between TRIB1High versus TRIB1Low MDMs and between TRIB1High (n = 191) versus TRIB1Low (n = 191) monocytes. (D) RT-qPCR quantification of RNA (mean ± SEM) in BMDM prepared from specified mice (n = 5 to 9 per group). (E) Immunocytochemistry of nonpolarized Trib1mWT and Trib1mTg BMDMs. OLR1 (red), nuclei counterstained with 4',6-diamidino-2-phenylindole (DAPI) (blue). Scale bar, 50 µm. Quantifications performed on BMDMs prepared from four to five mice per group. (F) Western blot analysis of OLR1 in Trib1mWT and Trib1mTg BMDMs (n = 3 to 5 per group). In (D) to (F), significance was determined by Student’s t test; *P < 0.05, and **P < 0.01.
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