Journal: Redox biology

Article Title: CD40-TRAF6 inhibition suppresses cardiovascular inflammation, oxidative stress and functional complications in a mouse model of arterial hypertension.

doi: 10.1016/j.redox.2025.103520

Figure Lengend Snippet: Fig. 1. CD40-TRAF6 inhibition leads to phenotypic improvement in hypertensive mice. (A) WT animals were treated with either TRAF6i (2.5 mg/kg/d), AT-II (1 mg/kg/d), or AT-II and TRAF6i via osmotic minipumps for seven days. The blood pressure was measured on day five, echocardiography was performed on day six, and the organs were harvested on day seven. (B) At the end of the treatment, the heart and body weight were measured to calculate the heart-to-body weight ratio. (C, D) The systolic blood pressure was determined on day six of the treatment via non-invasive tail-cuff blood pressure measurement. The systolic blood pressure data were reused from our previously published work [11] with permission, whereas diastolic blood pressure data were not shown before. (E) Endothelial function in response to acetylcholine was measured via isometric tension studies. (F) Aortic O2 −• production was analyzed by using the chemiluminescence probe lucigenin. (G, H) The chemiluminescence of L-012 oxidation was utilized to analyze the mainly leukocyte-dependent H2O2 production in whole blood samples. The blood was stimulated with PDBu for 10 min (G) or zymosan A for 55 min (H). Data are presented as mean values ± SD of n = 15–16 (B), n = 9–10 (C, D), n = 6–16 (E), n = 3 (F) and n = 5–12 (G, H) animals per group. *p ≤0.05, **p ≤0.01, and ***p ≤0.001. One-way ANOVA with Tukey’s multiple comparison test (B, C, D, F, G, H) and two-way ANOVA with Dunnett’s multiple comparison test (E) were performed. (A) and all icons were created with BioRender.com. Abbreviations: WT=C57BL6/J mice, TRAF6i = TRAF6 inhibitor, AT-II = angiotensin-II, bp = blood pressure, Ach = acetylcholine, PDBu = phorbol 12,13-dibutyrate.

Article Snippet: Male C57BL/6J (9–10 weeks old) mice were purchased from Charles River (Sulzfeld, Germany) and treated with either angiotensin-II (AT-II; 1 mg/kg/d in 0.9 % NaCl) [11,15], TRAF6 inhibitor (compound 6877002, Tocris, Bristol, UK; 2.5 mg/kg/d in 2.5 % DMSO) [4,11] or both compounds by osmotic minipumps (Alzet model 1007D, Cupertino, CA, USA) for seven days.

Techniques: Inhibition, Comparison

Journal: Redox biology

Article Title: CD40-TRAF6 inhibition suppresses cardiovascular inflammation, oxidative stress and functional complications in a mouse model of arterial hypertension.

doi: 10.1016/j.redox.2025.103520

Figure Lengend Snippet: Fig. 2. CD40-TRAF6 inhibition leads to reduced ROS formation in different tissues of hypertensive mice. The overall ROS production was analyzed in cryosections via DHE staining in different tissues: pVAT (A), heart (B), and kidney (C). Representative DHE pictures are shown below the bar graphs. The red color reflects the fluorescence from O2 −• and H2O2. Data are presented as mean values ± SD of n = 4–8 (A–C) animals per group. *p ≤0.05, **p ≤0.01, ***p ≤0.001, and ****p ≤0.0001. One-way ANOVA with Tukey’s multiple comparison test was performed. Icons were created with BioRender.com. Abbreviations: WT=C57BL6/J mice, TRAF6i = TRAF6 inhibitor, AT-II = angiotensin-II, DHE = dihydroethidium, pVAT = perivascular adipose tissue, Ao = aorta.

Article Snippet: Male C57BL/6J (9–10 weeks old) mice were purchased from Charles River (Sulzfeld, Germany) and treated with either angiotensin-II (AT-II; 1 mg/kg/d in 0.9 % NaCl) [11,15], TRAF6 inhibitor (compound 6877002, Tocris, Bristol, UK; 2.5 mg/kg/d in 2.5 % DMSO) [4,11] or both compounds by osmotic minipumps (Alzet model 1007D, Cupertino, CA, USA) for seven days.

Techniques: Inhibition, Staining, Fluorescence, Comparison

Journal: Redox biology

Article Title: CD40-TRAF6 inhibition suppresses cardiovascular inflammation, oxidative stress and functional complications in a mouse model of arterial hypertension.

doi: 10.1016/j.redox.2025.103520

Figure Lengend Snippet: Fig. 3. CD40-TRAF6 inhibition reduces vascular immune cell infiltration in hypertensive mice. The immune cell content in aortic tissue lysates was analyzed via flow cytometry with different fluorophore-tagged antibodies. The following antibodies were used to distinguish between different immune cell subtypes: (A) anti- CD45 (leukocytes), (B) anti-CD11b (myeloid cells), (C + D) anti-Ly6G and anti-Ly6C (inflammatory monocytes and neutrophils). Representative counter blots are shown below the bar graphs, and the area of interest is marked with a red box. Data are presented as mean values ± SD of n = 5–6 (A–D) animals per group. *p ≤ 0.05 and **p ≤0.01. One-way ANOVA with Tukey’s multiple comparison test was performed. Icons were created with BioRender.com. Abbreviations: WT=C57BL6/ J mice, TRAF6i = TRAF6 inhibitor, AT-II = angiotensin-II, CD = cluster of differentiation, Infl = inflammatory.

Article Snippet: Male C57BL/6J (9–10 weeks old) mice were purchased from Charles River (Sulzfeld, Germany) and treated with either angiotensin-II (AT-II; 1 mg/kg/d in 0.9 % NaCl) [11,15], TRAF6 inhibitor (compound 6877002, Tocris, Bristol, UK; 2.5 mg/kg/d in 2.5 % DMSO) [4,11] or both compounds by osmotic minipumps (Alzet model 1007D, Cupertino, CA, USA) for seven days.

Techniques: Inhibition, Flow Cytometry, Comparison

Journal: Redox biology

Article Title: CD40-TRAF6 inhibition suppresses cardiovascular inflammation, oxidative stress and functional complications in a mouse model of arterial hypertension.

doi: 10.1016/j.redox.2025.103520

Figure Lengend Snippet: Fig. 4. CD40-TRAF6 inhibition does not reduce left ventricular hypertrophy in hypertensive mice. After six days of treatment, transthoracic echocardiography for small animals was performed to determine left ventricular (LV) function. The Vevo Lab 3100 software was used to calculate the following parameters: (A) LV-EF, (B) LV mass, and (C) LV PW (systolic). Representative M-mode pictures are shown below the bar graphs. Data are presented as mean values ± SD of n = 12–16 (A–C) animals per group. **p ≤0.05, **p ≤0.01, ***p ≤0.001, and ****p ≤0.0001. One-way ANOVA with Tukey’s multiple comparison test was performed. Icons were created with BioRender.com. Abbreviations: WT=C57BL6/J mice, TRAF6i = TRAF6 inhibitor, AT-II = angiotensin-II, LV = left ventricle, EF = ejection fraction, PLAX = parasternal long axis view, M-mode = motion mode, PW = posterior wall, sys = systole, IVS = intraventricular septum, LVID = left ventricular internal diameter, LVPW = left ventricular posterior wall.

Article Snippet: Male C57BL/6J (9–10 weeks old) mice were purchased from Charles River (Sulzfeld, Germany) and treated with either angiotensin-II (AT-II; 1 mg/kg/d in 0.9 % NaCl) [11,15], TRAF6 inhibitor (compound 6877002, Tocris, Bristol, UK; 2.5 mg/kg/d in 2.5 % DMSO) [4,11] or both compounds by osmotic minipumps (Alzet model 1007D, Cupertino, CA, USA) for seven days.

Techniques: Inhibition, Software, Comparison

Journal: Redox biology

Article Title: CD40-TRAF6 inhibition suppresses cardiovascular inflammation, oxidative stress and functional complications in a mouse model of arterial hypertension.

doi: 10.1016/j.redox.2025.103520

Figure Lengend Snippet: Fig. 5. Adipocyte-specific CD40 knockout shows no vasoprotective phenotype in hypertensive mice. (A) Adipocyte-specific CD40−/− (AdiCD40−/−) and respective control mice (Ctr) were treated for seven days with AT-II (1 mg/kg/d) via osmotic minipumps. The blood pressure was measured on days five and six. The organs were harvested on day seven. (B) At the end of the treatment, the heart and body weight were measured to calculate the heart-to-body weight ratio. (C) The systolic blood pressure of the mice was determined via non-invasive tail-cuff blood pressure measurement. (D) Endothelial function in response to acetylcholine was measured via isometric tension studies. The ROS production was analyzed in cryosections via DHE staining in pVAT (E) and cardiac (F) tissue. Representative DHE pictures are shown below the bar graphs. Data are presented as mean values ± SD of n = 5–7 (B), n = 8–14 (C), n = 9–14 (D), and n = 4 (E, F) animals per group. *p ≤0.05, **p ≤0.01, ***p ≤0.001, and ****p ≤0.0001. One-way ANOVA with Tukey’s multiple comparison test (B, C, E, F) and two-way ANOVA with Dunnett’s multiple comparison test (D) were performed. Icons were created with BioRender.com. Abbreviations: Ctr = AdiCD40−/−(Cre WT), TRAF6i = TRAF6 inhibitor, AT-II = angiotensin-II, bp = blood pressure, DHE = dihydroethidium, pVAT = perivascular adipose tissue, ACh = acetylcholine.

Article Snippet: Male C57BL/6J (9–10 weeks old) mice were purchased from Charles River (Sulzfeld, Germany) and treated with either angiotensin-II (AT-II; 1 mg/kg/d in 0.9 % NaCl) [11,15], TRAF6 inhibitor (compound 6877002, Tocris, Bristol, UK; 2.5 mg/kg/d in 2.5 % DMSO) [4,11] or both compounds by osmotic minipumps (Alzet model 1007D, Cupertino, CA, USA) for seven days.

Techniques: Knock-Out, Control, Staining, Comparison