Journal: Oxidative Medicine and Cellular Longevity

Article Title: Anti-Interleukin-16 Neutralizing Antibody Treatment Alleviates Sepsis-Induced Cardiac Injury and Dysfunction via the Nuclear Factor Erythroid-2 Related Factor 2 Pathway in Mice

doi: 10.1155/2021/6616422

Figure Lengend Snippet: Establishment and pretreatment of the mouse sepsis model. WT: wild-type; PBS: phosphate-buffered saline; SOD: superoxide dismutase; LPS: lipopolysaccharide; CLP: cecal ligation and puncture; nAb: anti-IL-16 neutralizing antibody; CPU: CPUY192018; DMSO: dimethyl sulfoxide.

Article Snippet: Additionally, mice that received the anti-IL-16 nAb or IgG were also given 50 μ l of dimethyl sulfoxide (DMSO, Sigma) or 5 mg/kg CPUY192018 (Sigma), a nuclear factor erythroid-2 related factor 2 (Nrf2) pathway inhibitor, and were treated with LPS (part 5, n = 10) [ ].

Techniques: Ligation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Anti-Interleukin-16 Neutralizing Antibody Treatment Alleviates Sepsis-Induced Cardiac Injury and Dysfunction via the Nuclear Factor Erythroid-2 Related Factor 2 Pathway in Mice

doi: 10.1155/2021/6616422

Figure Lengend Snippet: Effects of CPUY192018 on cardiac injury in septic mice. (a) Survival rates were measured for each group (log-rank test). (b–d) Serum levels of cardiac injury markers, including CK-MB, LDH, and cTnT, were measured (two-way ANOVA). (e–h) Cardiac structure and function in each group were measured (two-way ANOVA). n = 5 in each group. ∗ p < 0.05 vs. the nAb DMSO+LPS group. LPS: lipopolysaccharide; DMSO: dimethyl sulfoxide; CPU: CPUY192018; nAb: anti-IL-16 neutralizing antibody; LDH: lactate dehydrogenase; CK-MB: creatine kinase myocardial bound; cTnT: cardiac troponin T; LVEDD: left ventricular end-diastolic diameter; LVESD: left ventricular end-systolic diameter; LVEF: left ventricular ejection fraction; FS: fractional shortening.

Article Snippet: Additionally, mice that received the anti-IL-16 nAb or IgG were also given 50 μ l of dimethyl sulfoxide (DMSO, Sigma) or 5 mg/kg CPUY192018 (Sigma), a nuclear factor erythroid-2 related factor 2 (Nrf2) pathway inhibitor, and were treated with LPS (part 5, n = 10) [ ].

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Anti-Interleukin-16 Neutralizing Antibody Treatment Alleviates Sepsis-Induced Cardiac Injury and Dysfunction via the Nuclear Factor Erythroid-2 Related Factor 2 Pathway in Mice

doi: 10.1155/2021/6616422

Figure Lengend Snippet: Effects of CPUY192018 on oxidative stress and cardiomyocyte apoptosis in septic mice. (a) Nrf2, HO-1, Nox2, Nox4, and GAPDH expression in LV tissue was detected (two-way ANOVA). (b) Serum activity of SOD and NADPH oxidase and levels of GSH and MDA were measured (two-way ANOVA). (c) AIF and Cox IV expression in mitochondria and AIF, cleaved PARP, and PCNA expression in nuclei were analyzed (two-way ANOVA). n = 5 in each group. ∗ p < 0.05 vs. the nAb DMSO+LPS group. Nrf2: nuclear factor erythroid-2 related factor 2; HO-1: heme oxygenase 1; Nox2/4: nicotinamide adenine nucleotide phosphate oxidase 2/4; nAb: anti-IL-16 neutralizing antibody; CPU: CPUY192018; LPS: lipopolysaccharide; SOD: superoxide dismutase; GSH: glutathione; NADPH: nicotinamide adenine nucleotide phosphate; MDA: malondialdehyde; AIF: apoptosis-inducing factor; Cox IV: cytochrome c oxidase IV; PARP: poly-ADP-ribose polymerase; PCNA: proliferating cell nuclear antigen.

Article Snippet: Additionally, mice that received the anti-IL-16 nAb or IgG were also given 50 μ l of dimethyl sulfoxide (DMSO, Sigma) or 5 mg/kg CPUY192018 (Sigma), a nuclear factor erythroid-2 related factor 2 (Nrf2) pathway inhibitor, and were treated with LPS (part 5, n = 10) [ ].

Techniques: Expressing, Activity Assay

Journal: Redox Biology

Article Title: A novel role of KEAP1/PGAM5 complex: ROS sensor for inducing mitophagy

doi: 10.1016/j.redox.2021.102186

Figure Lengend Snippet: Inhibitors of KEAP1–PGAM5 protein-protein interaction enhance mitophagy. A. Molecular docking simulation showing energetically most favorable interactions between a KEAP1 and non-electrophilic KEAP1 inhibitors. The table is showing hydrogen-bond pairings in KEAP1-inhibitor complexes. The length of the hydrogen bond is given in angstroms. B. Treatment with CPUY192018 disrupts KEAP1-PGAM5 interaction. PC6 cells were co-transfected with KEAP1-FLAG and PGAM5-HA and treated for 24 h with DMSO or 100 μM CPUY192018. KEAP1-FLAG was co-immunoprecipitating with PGAM5-HA in DMSO but not in CPUY192018 treated cells. C. Treatment with CPUY192018 increases the level of endogenous PGAM5. PC6 cells were treated with 100 μM CPUY192018 for 24 h. The left panel shows a representative Western blot and the right panel quantitative analysis. *P < 0.05, n = 4 samples, t -test. D. Effect of non-electrophilic KEAP1 inhibitors on mitochondrial membrane potential. PC6 cells were treated with KEAP1 inhibitors for 24 h, after which the cells were stained with ratiometric mitochondrial membrane potential sensor JC10. *P < 0.05 and ****P < 0.0001 when compared with DMSO treated group, n = 7–12 wells per data point, One-way ANOVA followed by Dunn's multiple comparisons test. E. Non-electrophilic KEAP1 inhibitors induce Parkin-EYFP translocation to mitochondria. PC6 cells expressing Parkin-EYFP were treated with DMSO or 100 μM of KEAP1 inhibitors for 24 h ***P < 0.001 and ****P < 0.0001 when compared with DMSO treated group, n = 6 dishes per group, 20 fields per dish, One-way ANOVA followed by Dunnett's multiple comparisons test. F and G. Non-electrophilic KEAP1 inhibitors enhance mitophagy but not general autophagy. Neurons expressing mitochondrial marker Kate2 and autophagosome markers EGFP-LC3B and GFP-LC3C were treated with DMSO or 100 μM of KEAP1 inhibitors for 24 h. The number of LC3 positive puncta colocalizing with mitochondria (F) as well as the total number of LC3 positive puncta (G) was counted by a blinded observer. **P < 0.01, ***P < 0.001 and ****P < 0.0001 when compared with DMSO treated group, n = 9–14 dishes (10 cells per dish), Welch's ANOVA followed by Dunnett's T3 multiple comparisons test. H. Treatment of primary cortical neurons with CPUY192018 (24 h) induces a concentration-dependent increase in mitochondrial colocalization with autophagosome markers EGFP-LC3B/GFP-LC3C. *P < 0.05 and ****P < 0.0001, when compared with DMSO, treated group, n = 16–18 dishes from 4 independent experiments (10 cells per dish), Kruskal-Wallis test followed by Dunn's multiple comparisons test, the dotted line shows the EC50.

Article Snippet: CPUY192018 , AOBIOUS, INC , Cat# AOB 9974.

Techniques: Transfection, Western Blot, Membrane, Staining, Translocation Assay, Expressing, Marker, Concentration Assay

Journal: Redox Biology

Article Title: A novel role of KEAP1/PGAM5 complex: ROS sensor for inducing mitophagy

doi: 10.1016/j.redox.2021.102186

Figure Lengend Snippet:

Article Snippet: CPUY192018 , AOBIOUS, INC , Cat# AOB 9974.

Techniques: Recombinant, Lysis, Extraction, Blocking Assay, Isolation, DC Protein Assay, Caspase-Glo Assay, shRNA, Generated, Plasmid Preparation, Synthesized, Variant Assay