Fig 1: The effect of UA on CCl4-induced liver injury and fibrosis is related to NOX4. (A) HE staining (100× magnification). (B) Masson’s trichrome staining (100× magnification). (C–D) Morphometrical analysis of the fibrotic score and fibrotic area. (E) Detection of the hydroxyproline content in the liver tissue by colorimetry. (F) Liver function indices in mouse sera. Data represent the mean ± SD for each group. *P < 0.05 and ***P < 0.001.
Fig 2: Autophagy regulated OS and senescence in PTCs of Aldo-induced rats. (A) Senescence-associated-galactosidase (SA-ß-Gal) Staining in rat renal cortex (n = 6). SA-ß-Gal is labeled by bright-blue in the PTCs. (B) Bar graph indicating the percentage of SA-ß-Gal positive cells per field in tubular. (C) DHE staining of rat kidney sections (n = 6). (D) Bar graph indicating the mean DHE intensity per field in rat tubular cells. (E) Immunohistochemical staining for NOX4 in rat kidney tissues from various groups, as indicated (n = 6). (F) Bar graph indicating NOX4 immunoreactivity per field in rat tubular cells. (G) Western blot analysis revealed the expression of NOX4, p21, and GAPDH proteins after various treatments in rats (n = 3). (H) Graphical presentation shows the relative abundance levels of NOX4 and p21 after normalization with GAPDH (n = 3). #p < 0.05 vs. normal control, *p < 0.05 vs. Aldo alone.
Fig 3: The IC50 curve shows the cytotoxicity assay of (a) HGC-27 and (d) MGC-803 cell lines treated with Erastin. The flow cytometry and fluorescence microscope plots verify the different ROS levels of (b, c) HGC-27 and (e, f) MGC-803 cell lines treated with Erastin. The boxplots indicate the different mRNA expression levels of these 10 FDEGs in (g) HGC-27 and (h) MGC-803 after being treated with 10 µM Erastin by real-time PCR. (i) The plot detects the different protein expression levels of hub FDEGs (SP1, KEAP1, NOX4, AIFM2, and GPX4) in HGC-27 and MGC-803 after being treated with 10 µM Erastin by western blot. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.
Fig 4: Functional role of Nox4 in the regulation of vascular dysfunction by docetaxel in vivo.(A) Schematic design of in vivo studies of the effects of docetaxel on vascular endpoints. Nox4–/– and WT mice were injected i.p. with docetaxel (10 mg/kg) or placebo (solvent) every 5 days for 3 weeks. (B) Effect of docetaxel on blood pressure (BP) in C57BL/6J mice by telemetry. Data are expressed as the mean ± SEM. *P < 0.05 versus WT; 2-way, repeated-measures ANOVA. (C) Systolic blood pressure by tail-cuff plethysmography in Nox4–/– and WT mice treated with docetaxel or placebo (n = 10–14/group). Data are expressed as the mean ± SEM. #P < 0.01 versus baseline; **P < 0.01 versus WT or docetaxel-treated Nox4–/– mice; 2-way, repeated-measures ANOVA with Tukey’s test. (D) Endothelium-dependent vasorelaxation to ACh (1 nM–10 µM) and endothelium-independent vasorelaxation to SNP (1 nM–10 µM) in mouse aortas (n = 9–14/group). Data are expressed as the mean ± SEM. ***P < 0.001 versus WT; *P < 0.05 versus Nox4–/– mice treated with docetaxel; 2-way, repeated-measures ANOVA with Tukey’s test. (E) H2O2 production using Amplex Red in aortas from Nox4–/– and WT mice treated with docetaxel or placebo (n = 9/group). Data are expressed as the mean ± SEM. **P < 0.01 versus Nox4–/– mice treated with docetaxel; *P < 0.05 versus WT; 2-way ANOVA with Bonferroni’s test. (F) LGCL (5 µM) in aortas (n = 8–9/group). Data are expressed as the mean ± SEM. **P < 0.01 versus WT; 2-way ANOVA with Bonferroni’s test. (G) Representative images showing superoxide production in mouse aortas using DHE fluorescence. Scale bars: 100 µm. (H) Nox4 mRNA expression in mouse aortas (n = 5/group). Data are expressed as the mean ± SEM. ****P < 0.0001 versus Nox4–/– mice treated with docetaxel; *P < 0.05 versus WT; 2-way ANOVA with Bonferroni’s test. (I) p-eNOS (Thr495) in Nox4–/– and WT mouse aortas treated with docetaxel or placebo (n = 5–6/group). Densitometric analysis was normalized to total eNos. Immunoblots represent 1 of 2 independent experiments and are summarized as the mean ± SEM. **P < 0.01 versus Nox4–/– mice treated with docetaxel; ***P < 0.001 versus WT; 2-way ANOVA and Bonferroni’s test.
Fig 5: Effect of luminal flow on Nox4 localization and activation in thick ascending limbs. Panels A and B: Effect of luminal flow on Nox4 localization in thick ascending limbs using an antibody against the carboxyl terminus of Nox4 (C-Nox4); (A) Representative immunofluorescent confocal images showing C-Nox4 staining (left panel) and same nuclei counterstained with POPO3 (right panel). (B) Quantification using the ratio between nuclear and cytoplasmic intensities (N/C intensity; n = 5 and 6, for no flow vs. flow, respectively). Panels C and D: Effect of luminal flow on Nox4 localization in thick ascending limbs using an antibody against the amino terminus of Nox4 (N-Nox4); (C) Representative immunofluorescent confocal images showing N-Nox4 staining (left panel) and same nuclei counterstained with POPO3 (right panel). (D) Quantification using the ratio between nuclear and cytoplasmic intensities (N/C intensity; n = 6 for each group). Nuclei are outlined for better identification.
Supplier Page from Abcam for Anti-NADPH oxidase 4 antibody [UOTR1B492]