Fig. 2

NOX4 stimulates FOXM1 expression by increasing mitochondrial ROS. A Analysis of NOX4 expression in normal brain and glioma tissues in TCGA datasets. B The prognostic value of NOX4 expression in LGG and GBM was analyzed in TCGA datasets. C qPCR detection of NOX4 mRNA expression in normal brain and glioma clinical specimens. D IHC analysis of NOX4 protein expression in normal brain and GBM. Scale bars, 20 μm. E Western blot analysis of NOX4 and FOXM1 protein expression in normal brain and glioma tissues of different grades. F The correlation between NOX4 expression and FOXM1 expression in normal brain and glioma patients according to the TCGA and our clinical samples. G Western blot and qPCR analysis of FOXM1 protein and mRNA levels in control or NOX4-overexpressing glioblastoma cells. H MitoSOX staining showing the upregulation of NOX4 affected the production of ROS in glioblastoma cells. Scale bars, 50 μm. I Western blot and qPCR analysis of FOXM1 protein and mRNA levels in control or NOX4 knockdown glioblastoma cells. J MitoSOX staining showed that the downregulation of NOX4 affected the production of ROS in glioblastoma cells. Scale bars, 50 μm. K Western blot analysis showing the protein expression of FOXM1 in glioblastoma cells exposed to NAC at different concentrations for 24 h. L Western blot analysis showing the protein expression of FOXM1 in glioblastoma cells exposed to H₂O₂ at different concentrations for 24 h. M Western blot analysis showing the protein expression of FOXM1 in glioblastoma cells treated with Mito-Tempo (1 mM) in the control or NOX4-overexpressing cells. N Immunofluorescence staining showing the subcellular localization of NOX4 in glioblastoma cells; the merge in yellow shows colocalization. Scale bar 50 μM. *, P < 0.05; **, P < 0.01; ***, P < 0.001