Fig. 5

CPO inhibits HUVEC proliferation through a copper–mitochondria regulatory mechanism.(A) Representative western blot showing the expression of DLAT, SDHB and HSP70 in different groups normalized to the expression of β-actin. (B) Densitometric analyses of DLAT, SDHB and HSP70 expression in different groups normalized to the expression of β-actin. (C) Immunofluorescence staining showing DLAT (green) and VDAC1 (red). White scale bar = 20 μm. (D) Quantified immunostaining of DLAT foci in different groups. (E) TEM results showing the mitochondrial structure in the different groups. The white arrows indicate normal mitochondria. The orange arrows indicate swelling. The yellow arrows indicate vacuoles. Scale bar = 1.0 μm. Yellow scale bar = 0.25 μm. (F) Quantification of the mitochondrial number, abnormal mitochondrial percentage, and average mitochondrial minor axis length. (G) JC-10 results showing the mitochondrial function of HUVECs in the different groups. (H) Column chart showing the ratio of aggregated JC-10 to monomeric JC-10 in the different groups. The data are presented as the means ± SDs of three independent experiments. The error bars represent the ± SDs. ns: P > 0.05, *: P < 0.05, **: P < 0.01. CPO: copper overload; HUVEC: human umbilical vein endothelial cell; ES: elesclomol; TEM: transmission electron microscopy; SD: standard deviation