Fig. 4

TAX1BP1 was the major receptor of CuONPs-induced mitophagy in vascular endothelial cells. A MTS analysis was performed to detect EA.hy926 cell viability. Cells were transfected with siNC, siGFP, siTAX1BP1, siNBR1, sip62, siOPTN or siNDP52 for 48 h, and then treated with CuONPs (0, 5, 7.5, 10 and 15 μg/ml) for 24 h. B Representative immunofluorescence images indicates colocalization of TAX1BP1 with GFP-LC3 and Mito-DsRed dots in EA.hy926 stable cell line expressing GFP-LC3 and Mito-DsRed after treatment with 10 μg/ml CuONPs for 12 h. White arrow indicate TAX1BP1-positive dots that colocalized with GFP-LC3 and Mito-DsRed signals. Scale bars, 20 μm. C and D Representative FCM results of EA.hy926 cells labeled with MitoTracker Green FM (C) and MitoSOX (D), respectively. The cells were transfected with siNC or siTAX1BP1 for 48 h and then treated with CuONPs (10 μg/ml) for 12 h. All data were statistically analyzed using one-way ANOVA with Tukey’s test. Results are representative of at least three independent experiments. Data are mean ± S.D. *p < 0.05