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. 2018 May 22;175(13):2581–2598. doi: 10.1111/bph.14174

Figure 4.

Figure 4

NOX4 mediates 4‐HNE‐induced superoxide production and cell death in ARPE‐19 cells. 4‐HNE induced ROS production, which was quenched by (A) pretreatment with SOD, (B) siRNA knockdown of GPR109A, GPR109B or NOX4, (C) treatment with inhibitors of Gβγ‐downstream molecules and (D) treatment NADPH oxidase inhibitors. * P < 0.05 versus vehicle‐treated or non‐targeting siRNA‐transfected controls, # P < 0.05 versus cells treated with 10 or 30 μM 4‐HNE. (E) Copy numbers of the NADPH oxidase subunits NOX1, NOX2 or NOX4 in ARPE‐19 cells were measured by qRT‐PCR. * P < 0.05 versus vehicle‐treated controls. (F) Western blotting results for NOX4 induction in 4‐HNE‐treated cells. * P < 0.05 versus vehicle‐treated controls. (G) Caspase‐3/7 activity was measured in ARPE‐19 cells pretreated with NOX4 siRNA prior to 4‐HNE treatment. * P < 0.05 versus vehicle‐treated controls. # P < 0.05 versus non‐targeting siRNA‐transfected cells. (H) Cell viability measurements in cells treated with 4‐HNE in the presence of NOX4 and mitochondrial inhibitors or antioxidants. * P < 0.05 versus vehicle‐treated controls. # P < 0.05 versus 4‐HNE‐treated cells. $ P < 0.05 versus 4‐HNE plus antimycin A‐treated cells.