Fig. 5

UA alleviated CuONPs-induced cell death. A Representative fluorescence images of EA.hy926 cell line stable expressing GFP-LC3 and Mito-DsRed. B Representative fluorescence images of EA.hy926 cell line stable expressing PINK1-GFP and Mito-DsRed. In A and B, the cells were treated with UA (100 μM), CuONPs (10 μg/ml) and UA + CuONPs for 12 h, respectively. White arrows indicate PINK1-positive or LC3-positive dots that colocalized with damaged mitochondria signals. Scale bars, 20 μm. C Representative FCM results of EA.hy926 staining with mitochondrial mass marker MitoTracker Green FM. MFI, mean fluorescence intensity. D Western blotting analysis and quantification of protein levels of TOM20, VDAC1, ATP5B and TIM23 in CuONPs-treated EA.hy926 cells with or without UA. E Representative FCM results of EA.hy926 staining with mtROS probe MitoSOX. F Representative FCM results of EA.hy926 staining with 7-AAD. EA.hy926 cells were transfected with siNC or siPINK1 for 48 h, and then treated with UA (100 μM) and CuONPs (10 μg/ml) for 24 h. n.s., not significance. The cells were treated with CuONPs (10 μg/ml) or UA + CuONPs for 12 h, respectively. All data were statistically analyzed using One-way ANOVA with Tukey’s test. The results are representative of at least three independent experiments. Data are mean ± S.D. *p < 0.05