Fig. 3

NRF2 protects against CuONPs-triggered oxidative stress and cell death in HUVECs. A Representative images of cell morphology of WT and NRF2-KO cells treated with 20 μg/ml CuONPs for 12 h. Scale bar, 100 μm. B, C Immunoblotting analysis and quantification of protein levels of NRF2, HMOX1, γH2AX and β-Actin (loading control) in WT and NRF2-KO cells treated with 20 μg/mL CuONPs for 0, 6 and 9 h, respectively. D, E FACS analysis and quantification of DHE intensity in WT, HMOX1 knockout (HMOX1-KO) or NRF2-KO cells treated with CuONPs (20 μg/ml) for 12 h, respectively. Unstained, non-labeled cells. MFI, mean fluorescence intensity. E, F FACS analysis and quantification of 7-AAD intensity in WT, HMOX1-KO and NRF2-KO cells treated with CuONPs (20 μg/ml) for 12 h, respectively. H, I FACS analysis and quantification of DHE intensity in WT and NRF2-KO cells treated with NAC (10 mM) for 1 h before treatment with CuONPs (20 μg/ml) for 12 h, respectively. J, K FACS analysis and quantification of 7-AAD intensity in WT and NRF2-KO cells treated with NAC and CuONPs. All data are representative of three independent experiments. Statistical significance was evaluated using one-way ANOVA followed by a Tukey multiple comparison test. The values are expressed in mean ± S.D. ns not significance; “***”, “P ≤ 0.001”