Journal: Biochemistry and Biophysics Reports

Article Title: The miR-2110/ TRAF3 axis is associated with endothelial dysfunction and atherosclerosis in coronary heart disease

doi: 10.1016/j.bbrep.2026.102508

Figure Lengend Snippet: Validation of TRAF3 as a direct target of miR-2110 by RT-qPCR, luciferase reporter assay, and Western blot analysis ( A ) RT-qPCR analysis of TRAF3 mRNA levels in OE2110 and OENC cells, calculated using the 2 −ΔΔCt method, normalized to GAPDH, and expressed relative to the control group (set to 1). ( B ) TRAF3 mRNA levels in peripheral blood samples from healthy controls (n = 30) and CHD patients (n = 34), normalized to GAPDH. ( C ) Schematic representation of the predicted binding sites between miR-2110 and the wild-type (WT) 3′-UTR of TRAF3 , and the corresponding mutated (MT) sequence used for luciferase assays. ( D ) Dual-luciferase reporter assay showing decreased luciferase activity in cells co-transfected with miR-2110 mimics and TRAF3 -WT but not TRAF3 -MT. ( E, F ) Western blot analysis and quantification of TRAF3 protein expression in OE2110 and OENC, with GAPDH as internal reference. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ns: not significant. Panels A, D, and F: Data are presented as mean ± SD from three independent biological experiments (N = 3) using EA.hy926 cells. Panel E: Representative blots are shown. Statistical comparisons were performed using unpaired two-tailed Student's t-tests. Panel B (patient samples): Data were presented as median (interquartile range) and were analyzed using the Mann–Whitney U test.

Article Snippet: Endogenous peroxidase activity was quenched with 3% H 2 O 2 for 10 min at room temperature, followed by blocking with 5% normal goat serum (Solarbio, China, SL038) in PBS for 1 h. Sections were incubated overnight at 4 °C with rabbit anti-TRAF3 monoclonal antibody (1:200, HUABIO, China, PSH01-30), then probed with HRP-conjugated goat anti-rabbit IgG (1:500, Servicebio, China, GB23303) for 1 h at room temperature.

Techniques: Biomarker Discovery, Quantitative RT-PCR, Luciferase, Reporter Assay, Western Blot, Control, Binding Assay, Sequencing, Activity Assay, Transfection, Expressing, Two Tailed Test, MANN-WHITNEY

Journal: Biochemistry and Biophysics Reports

Article Title: The miR-2110/ TRAF3 axis is associated with endothelial dysfunction and atherosclerosis in coronary heart disease

doi: 10.1016/j.bbrep.2026.102508

Figure Lengend Snippet: Increased TRAF3 expression in cardiac and aortic tissues of ApoE −/− mice fed a high-fat diet ( A ) Relative Traf3 mRNA expression in the aortic root and adjacent heart tissue quantified by RT-qPCR, calculated using the 2 −ΔΔCt method, normalized to GAPDH, and expressed relative to the control group (set to 1). ( B, C ) Western blot analysis and quantification of TRAF3 protein expression in heart tissues. ( D, E ) Western blot analysis and quantification of TRAF3 protein expression in aortic tissues. ( F ) Representative immunohistochemical staining of TRAF3 in aortic root sections from control and ApoE −/− HFD mice, showing predominant localization in the tunica intima, including the endothelial layer and subendothelial regions containing vascular smooth muscle cells and infiltrating leukocytes (200 × ). Red boxes highlight regions with enhanced staining in HFD mice. (G) Quantification of TRAF3-positive area in the tunica intima. ∗ P < 0.05. Data represent mean ± SD; n = 6 mice per group. Statistical comparisons were performed using unpaired two-tailed Student's t-tests.

Article Snippet: Endogenous peroxidase activity was quenched with 3% H 2 O 2 for 10 min at room temperature, followed by blocking with 5% normal goat serum (Solarbio, China, SL038) in PBS for 1 h. Sections were incubated overnight at 4 °C with rabbit anti-TRAF3 monoclonal antibody (1:200, HUABIO, China, PSH01-30), then probed with HRP-conjugated goat anti-rabbit IgG (1:500, Servicebio, China, GB23303) for 1 h at room temperature.

Techniques: Expressing, Quantitative RT-PCR, Control, Western Blot, Immunohistochemical staining, Staining, Two Tailed Test