Fig. 6

NOX4 regulates the expression of EZH2 and p-EZH2 through the canonical Wnt/β-catenin pathway. a NP cells were transfected with NOX4 vector for NOX4 overexpression and immunostained with antibodies for NOX4 (red) or EZH2 (green). The nuclei were stained with DAPI (blue). Scale bar, 25 μm. b The 18 genes which changed significantly in PCR array analysis (n = 3). The original PCR array analysis data are presented in Additional file 4: Table S1. c RT-qPCR analysis of EZH2 in NP cells overexpressing NOX4. The data are represented as the mean ± SEM (n = 3). d Reactive oxygen species (ROS) levels measured using the DCFH-DA ROS-sensitive dye and flow cytometry. e Immunoblot analysis for EZH2, p-EZH2 and β-catenin in cells treated with NOX4 inhibitor GKT137831 (20 μM, 24 h). f Immunoblot analysis for NOX4, β-catenin, Wnt6, Wnt11, Wif1, and Mapk8 in cells overexpressing NOX4. β-actin was used as a loading control. NP cells transfected with empty lentivirus vectors were used as a control. The data are represented as the mean ± SEM (n = 3). g Immunoblot analysis for NOX4, β-catenin, EZH2, and p-EZH2 in NP cells treated with the Wnt signaling pathway inhibitor KYA1797K (25 μM for 24 h), NOX4 vector, or both. β-actin was used as a loading control. DMSO was used as a control for the inhibitor. NP cells transfected with empty lentivirus vectors were used as controls for the groups overexpressing NOX4. The data are represented as the mean ± SEM (n = 3). *p < 0.05, **p < 0.01. Wnt6 Wnt family member 6, Wnt11 Wnt family member 11, Wif1 Wnt inhibitory factor 1, Mapk8 mitogen-activated protein kinase 8