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. 2013 Aug 14;8(8):e71359. doi: 10.1371/journal.pone.0071359

Figure 5. PI3K/Akt pathway mediates stretch-induced phosphorylation of eNOS and NO production in the late phase.

Figure 5

(A) Upper: Western blots of phospho-Akt (Ser473), phospho-eNOS (Ser1177), and phospho-PKA substrates in HUVECs under continuous stretch (15 min), pretreated with LY294002 (10–100 µM). Lower: quantitative analysis of Ser1177 phosphorylation of eNOS. (B) Western blots of phospho-Akt (Ser473), phospho-eNOS (Ser1177), and PKA substrates in HUVECs under continuous stretch (30 min), pretreated with LY294002 (10–100 µM). Lower: quantitative analysis of Ser1177 phosphorylation of eNOS. (C) Upper: Western blots of phospho-Akt (Ser473), phospho-eNOS (Ser1177), and PKA substrates in HUVECs expressing scrambled (Scr) or Akt1/2-targeting (Akt KD) shRNAs under continuous stretch. Lower: quantitative analysis of Ser1177 phosphorylation of eNOS. (D) Upper: DAF-FM staining of HUVECs expressing scrambled (Scr) or Akt1/2-targeting (Akt KD) shRNAs with or without 50 µM LY294002 pretreatment, under continuous stretch Lower: quantitative analysis of NO release. Results are representative of 3 individual experiments and expressed as mean ± S.D. (n = 4). *p<0.05; **p<0.01; N.S., not significant.