Fig. 1

The activation of NRF2 in CuONPs-treated HUVECs. A Representative confocal images of NRF2 in HUVECs cells treated with CuONPs (20 μg/mL) for 12 h. Scale bar, 20 μm. Nuclei were stained with DAPI. MG132 or arsenite were used as positive controls. B, C Immunoblotting analysis and quantification of protein levels of NRF2 and its downstream HMOX1 and GCLM in HUVECs cells treated with 0, 5, 10, 15 and 20 μg/mL CuONPs for 12 h, respectively. GAPDH served as the internal control. D, E Immunoblotting analysis and quantification of protein levels of NRF2, HMOX1 and GCLM in HUVECs cells treated with 20 μg/mL CuONPs for 0, 3, 6, 9 and 12 h, respectively. GAPDH served as the internal control. F qPCR analysis of the mRNA levels of HMOX1, GCLM, SLC7A11, NQO1 and TXN in wild-type (WT) or NRF2 knockout (NRF2-KO) cells treated with 20 μg/mL CuONPs for 0, 6 and 9 h, respectively. G, H Immunoblotting analysis and quantification of NRF2 and HMOX1 protein levels in WT or NRF2-KO cells treated with 20 μg/mL CuONPs for 0, 6 and 9 h, respectively. β-Actin was used as loading control. All data are representative of three independent experiments. In C and E, Student’s t-test was used for statistical analysis. In F and H, one-way ANOVA followed by a Tukey multiple comparison test was used for statistical analysis. The values are expressed in mean ± S.D. ns not significance; **”, P ≤ 0.01; “***”, “P ≤ 0.001”