Fig. 5: Overexpression of VPS35 rescued copper overload and cuproptosis caused by norepinephrine (NE). | Nature Communications

Fig. 5: Overexpression of VPS35 rescued copper overload and cuproptosis caused by norepinephrine (NE).

From: Sleep fragmentation exacerbates myocardial ischemia‒reperfusion injury by promoting copper overload in cardiomyocytes

Fig. 5

A Validation of the expression levels of VPS35. Statistical analysis of VPS35 expression (n = 3, independent experiments for B; n = 5 mice for C). D Validation of the overexpression of VPS35 and Flag in HL-1 cells 96 h after transfection with the lentivirus. E Statistical analysis of VPS35 overexpression (OE) (n = 3 independent experiments, P = 0.0091). F Representative images of copper overload and ATP7A localization in both wide-field and confocal immunofluorescence images by confocal microscopy (red: CS1, green: ATP7A, blue: DAPI). G CS1 staining was quantified by calculating the mean fluorescence intensity (n = 8 independent experiments). H Validation of the levels of DLAT oligomerization, lipoylated proteins, iron-sulfur cluster proteins and HSP70 by Western blotting after treating vector- and VPS35-overexpressing HL-1 cells with 10 µM CuCl2 and 10 µM NE for 24 h. I–N Statistical analysis of the levels of DLAT oligomers, DLAT monomer, Lip-DLAT, Lip-DLST, FDX1 and HSP70. The data on Lip-DLAT and Lip-DLST were normalized to tubulin, and the other data were normalized to GAPDH (n = 4 independent experiments). O Vector- and VPS35-overexpressing HL-1 cells were treated with 10 µM CuCl2 in the presence of different concentrations of NE for 12 h, and cell viability was determined using a CCK-8 assay (n = 6 independent experiments). All data are presented as the mean ± s.e.m. The significance of differences was evaluated using one-way ANOVA for B, G and I–N, unpaired two-tailed t-test for C and E, and two-way ANOVA for O. Source data are provided as a Source Data file.

Back to article page