Figure 3

Figure 3 HEK293 cell lines overexpressing the rare variants found in NOX4 increased NOX4 expression and reactive oxygen species (ROS) generation compared to cells overexpressing NOX4^wt.

(A) The expression of NOX4 was analyzed by qRT-PCR of cells extracted from HEK293 stable transfected cells expressing pcDNA3.1 (empty vector), NOX4^wt (wild-type), NOX4^Y512IfsX20, NOX4^Y512C, and NOX4^V369F. β-actin levels were used as a loading control. Relative levels of NOX4 were quantified from three independent experiments. N = 3. (B) All NOX4 rare variants expressing cells have a higher generation of ROS compared to NOX4^wt. N = 4. (C) The generation of H₂O₂ was assessed using the 1 mM BioTracker Green H₂O₂ live-cell dye for 20 min in HEK293 stable transfected cells. The mean fluorescence intensity of H₂O₂ was quantified using ImageJ. N = 4. (D) Representative immunofluorescence images of H₂O₂ levels in HEK293 cells expressing NOX4 rare variants. Scale bar: 100 μm. Data information: H₂O₂ hydrogen peroxide. N =  biological replicates. For graphs (A–C), data are shown as mean ± SEM based on the ordinary one-way ANOVA compared to NOX4^wt. Source data are available online for this figure.