Fig 1: The protein expression of NOX4 and ratio of p-p38 MAPK/p38 MAPK are upregulated in coronary artery tissues of AS mice. (A) The protein expressions of NOX4, p38 MAPK and p-p38 MAPK were detected by Western blot; (B) Statistical comparison of NOX4, p38 MAPK and p-p38 MAPK protein expressions among four groups; The data were analyzed by one-way ANOVA with Tukey's test; n = 10; * p < 0.05 vs. the normal group; # p < 0.05 vs. the AS group; AS, atherosclerosis; NOX4, NADPH oxidase 4.
Fig 2: A schematic for the function and mechanism of miR-363-3p in CHD. miR-363-3p targets NOX4 and disrupts the p38 MAPK/ signaling pathway, which consequently reduces the apoptosis of CAECs, oxidative stress injury and the expression of inflammatory factors in CAECs, thus protecting CAECs against CHD.
Fig 3: miR-363-3p reduces H2O2 level and CAT activity in CAECs by targeting NOX4. (A) quantification of H2O2 level in CAECs; (B) quantification of CAT activity in CAECs; the data were analyzed by one-way ANOVA with Tukey's post hoc test; n = 3; * p < 0.05 vs. the blank group; # p < 0.05 vs. the miR-363-3p mimic group; CAECs, coronary artery endothelial cells; CAT, catalase; miR, microRNA; NOX4, NADPH oxidase 4; NC, negative control.
Fig 4: Treatment of TGF-ß activates the NLRP3 inflammasome activation signals in LX-2 cells. a ROS formation was analyzed by H2-DCFDA assay. LX-2 cells were seeded in 96-well black plates and treated with 10 ng/ml TGF-ß for 10, 30, or 60 min. After treatment with TGF-ß1, the cells were treated with 10 uM H2-DCFDA for 20 min. Fluorescence was measured using a microplate reader. b LX-2 cells were treated as described in Fig. 2a, and protein levels of p-Smad2/3, smad2/3, NOX4, and NEK7 were analyzed by western blotting. GAPDH was used as an internal control. c Densitometric quantification results of western blot images are shown. Data are means ± SEM. **p < 0.01, *p < 0.05 vs. CON; ##p < 0.01, #p < 0.05 vs. TGF-ß (10 min); $$p < 0.01, $p < 0.05 vs. TGF-ß (30 min) (n = 3/group)
Fig 5: Reduced miR-363-3p expression and abundant NOX4 and p38 MAPK expressions in HCY-induced CAECs. (A) positive and negative staining for goat antibody to CD34 (scale bar = 25 µm); (B) positive and negative staining for rabbit antibody to Factor VIII (scale bar = 25 µm); (C) miR-363-3p expression and NOX4 mRNA expression were determined by RT-qPCR in HCY-induced CAECs; (D) Representative Western blots of NOX4, p38 MAPK and p-p38 MAPK proteins and their quantitation in HCY-induced CAECs, normalized by ß-actin. The data were analyzed by paired t-test. n = 3. * p < 0.05 vs. the control group; CAECs, coronary arterial endothelial cells; SP, streptavidin-peroxidase.
Supplier Page from Abcam for Anti-NADPH oxidase 4 antibody [UOTR1B492] (HRP)