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. 2017 Jan 13;13(2):404–422. doi: 10.1080/15548627.2016.1263776

Figure 5.

Figure 5.

Inhibited mitochondrial fusion impairs endothelial function. (A) HUVECs were transfected with control (siCtrl) or MFN2 siRNA (siMFN2) for 48 h, and mitochondria were labeled with MitoTracker® Deep Red FM. Mitochondrial morphology was analyzed using fluorescence microscopy. Scale bars: 5 µm. (B) Mitochondrial fragmentation was determined, as described in Materials and Methods. n ≥ 100; *P < 0.05 vs. siCtrl. (C) HUVECs were transfected with siCtrl or siMFN2 for 48 h and then treated with A23187 or DMSO for 30 min. Phosphorylation of NOS-3 was measured by western blotting. (D) Nitric oxide concentration in the cells was assessed using the DAF-2 fluorescence probe. *P < 0.05 vs. DMSO; #P < 0.05 vs. siCtrl. (E) WT mouse carotid arteries were transfected with siCtrl or siMfn2 using electroporation for 48 h, stained with antibodies against MFN2 (Red) and PECAM1 (Green), and observed using fluorescence microscopy. (F) Endothelium-dependent vasodilator responses were measured in the presence of Ach (10−9 to 10−5 M). (G) Endothelium-independent vasodilator responses were measured in the presence of SNP (10−10 to 10−6 M). n = 4, * P < 0.05 vs. siCtrl.