Fig. 1.

A and B: human aortic endothelial cells (HAEC) exposed to oxidized LDL (OxLDL, 0, 5, 10, 15, 25, and 50 µg/ml) for 24 h were subjected to Western blotting with cystathionine γ-lyase (CSEγ) and GAPDH antibodies. C: mRNA levels of CSEγ and 18s (housekeeping gene) were assessed by real-time PCR in HAEC exposed to OxLDL (50 µg/ml) for 24 h and presented as 2^–ΔΔCt (cycle threshold). D and E: isolated aortas from apolipoprotein E (ApoE) knockout (KO) mice fed a normal diet (ND) or a high-fat diet (HFD) for 12 wk were subjected to Western blotting with CSEγ and β-tubulin antibodies. IB, immunoblot. F–H: mouse aortas (F and G) and HAEC (H) were transduced with CSEγ adenoviruses (Ad) for 24 h, and cell lysates were subjected to Western blotting with total (t) endothelial nitric oxide (NO) synthase (eNOS), total Akt, phosphorylated (Ser¹¹⁷⁷) eNOS (p-eNOS), phosphorylated (Thr³⁰⁸) Akt (p-Akt), and GAPDH antibodies. I: electrochemical probes were used to assay cell lysates from HAEC transduced with CSEγ adenoviral construct [30 multiplicity of infection (MOI)] for H₂S content. J: standard curve for H₂S measurements by electrochemical probes. K: NO [nitrite (NOx)] release from HAEC transduced with increasing amounts of adenoviruses encoding CSEγ (0–100 MOI) measured using Siever’s NO analyzer. L: cell lysates from HAEC transduced with increasing amounts of adenoviruses encoding CSEγ (0–100 MOI) were immunoblotted with anti-CSEγ antibodies. *P < 0.05 vs. respective controls. Western blots are representative of 3 immunoblots.