Figure 7. Mito-Esc but not parent esculetin rescues oxidant-induced deregulation of Oxygen Consumption Rate (OCR) in endothelial cells.

(a) HAEC were treated with Mito-Esc, Esculetin and TPP⁺ as indicated for 6 h and oxygen consumption rate (OCR) was measured by using XF 24 Extracellular Flux Analyzer (Seahorse Bioscience). (b–f) Endothelial cells were pretreated with Mito-Esc (2.5 μM) for 2 h before H₂O₂ (0.5 mM) was added for another 4 h- (b) oxygen consumption rate (OCR) was measured by XF 24 Extracellular Flux Analyzer. Oligomycin (1 μM), FCCP (1 μM), Rotenone (1 μM) + Antimycin A (1 μM) were sequentially added (indicated by arrows) and measured the basal respiration (c) maximal respiration (d) Spare respiratory capacity (e) and ATP production (f). (g) Cells were pretreated with Mito-Esc (2.5 μM) for 2 h before H₂O₂ (0.5 mM) was added for another 4 h and the levels of OXPHOS complex subunits were measured by immunoblotting. Results presented are average of three independent experiments. *Significantly different (p < 0.05) compared to untreated conditions. ^#Significantly different (p < 0.05) compared H₂O₂ treated condition.