Figure 3. Mito-Esc restores oxidant-induced depletion of GSH levels, dysregulation of mitochondrial transmembrane potential and mitochondrial superoxide generation in endothelial cells.

(a) HAECs were pretreated with Mito-Esc (2.5 μM) or esculetin (2.5 μM) or TPP⁺ (2.5 μM) for 2 h before the addition of Ang-II (0.5 μM) for 24 h and H₂O₂ levels were measured by the Amplex red assay. (b) Cells were pretreated with Mito-Esc or esculetin for 8 h, and GSH levels were measured as described in methods section. (c) HAEC were pretreated with Mito-Esc for 2 h before H₂O₂ (500 μM) was added for 8 h, and mitochondrial transmembrane potential was measured using TMRE fluorescence as described in methods. (d) Cells were treated with H₂O₂ or Ang-II in the presence or absence of either Mito-Esc or esculetin or TPP⁺ for 8 h and mitochondrial superoxide generation was measured by Mito-SOX dye as described in Methods section. *Significantly different (p < 0.05) compared to untreated conditions. ^#Significantly different (p < 0.05) compared to Ang-II or H₂O₂ treated condition. ^$Significantly different (p < 0.05) compared to Esculetin + H₂O₂ treated condition. Results indicated were average of three independent experiments.