Figure 3. AMPKα and ERK pathways are not involved in stretch-induced phosphorylation of eNOS.

(A) Upper: Western blots of phospho-eNOS (Ser¹¹⁷⁷) and phospho-AMPKα (Thr¹⁷²) in HUVECs under continuous stretch, pretreated with Compound C (5–50 µM). Lower: Quantitative analysis of Ser¹¹⁷⁷ phosphorylation of eNOS. (B) Upper: Western blots of phospho-AMPKα (Thr¹⁷²) and phospho-eNOS (Ser¹¹⁷⁷) in HUVECs expressing scrambled (Scr) or AMPKα1-targeting (AMPKα1 KD) shRNAs under continuous stretch. Lower: Quantitative analysis of Ser¹¹⁷⁷ phosphorylation of eNOS. (C) Upper: Western blots of phospho-Erk1/2 (Thr²⁰²/Tyr²⁰⁴) and phospho-eNOS (Ser¹¹⁷⁷) in HUVECs after stretch, pretreated with PD98059 (5–50 µM). Lower: Quantitative analysis of Ser¹¹⁷⁷ phosphorylation of eNOS. (D) Quantitative analysis of NO release in HUVECspretreated with 10 µM Compound C or PD980598, under continuous stretch. Results are representative of 3 individual experiments and expressed as mean ± S.D. (n = 4). N.S., not significant.