Fig. 7: Modulation of KLF2 or Nox2 expression restored EDD function in endothelial mTORC1- or mTORC2-inhibited mice.

a, b EC were isolated from Rptor^EC−/− or Rictor^EC−/− and their WT littermates followed by western blotting. c EC isolated from wild-type mice were infected with empty AAV (AAV-GFP) or Klf2-expressing adeno-associated virus (AAV-Klf2) at a multiplicity of infection (MOI) of 10⁵vg/cell for 72 h, then treated with or without 1 nM rapa for 1 h followed by western blot analysis. d Murine EC was infected with AAV containing two tandem mouse Nox2-targeting sequences (AAV-shNox2) followed by western blot analysis. e, f Rptor^EC−/− mice were i.v. injected with AAV-GFP or AAV-Klf2 at 3 × 10¹¹vg. EDD and EID were evaluated with transcutaneous ultrasound imaging (n = 5, e), and serum NO evaluated by the colorimetric measurement (n = 5, f). g–i, Rictor^EC−/− mice were injected with 3 × 10¹¹vg AAV-GFP or AAV-shNox2 followed by evaluation of EDD and EID (n = 5, g), and evaluation of serum superoxide (n = 5, h) and NO by the colorimetric measurement (n = 4, i). Error bars correspond to the standard error of the mean (SEM). *p < 0.05; **p < 0.01 vs. AAV-GFP; unpaired two-tailed t test (e–g, i) or one sample t test (h).