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. 2017 Sep 6;7:10602. doi: 10.1038/s41598-017-10894-x

Figure 3.

Figure 3

IL-33 stimulates NO production in LECs via the PI3K/Akt/eNOS signalling pathway. (A–C) IL-33-induced (20 ng/mL) Akt and eNOS phosphorylation was determined by Western blotting. (A) HDLECs were stimulated with various concentrations of IL-33 for 30 min. (B) HDLECs were stimulated with IL-33 (20 ng/mL) for the indicated times. (C) HDLECs were pre-treated with wortmannin (100 nmol/L, 30 min) and then stimulated with IL-33 (20 ng/mL, 30 min). (D) HDLECs were pre-treated with wortmannin (100 nmol/L, 30 min) or NMA (1 mmol/L) and then treated with IL-33 (20 ng/mL) for 4 hours. LECs were incubated with DAF-FM DA for 1 hour at 37 °C. The relative intracellular NO levels were determined from the fluorescence intensity of DAF-FM. Three independent experiments were performed in duplicate. *p < 0.05, **p < 0.01, ***p < 0.001. Full-length blots are presented in Supplementary Figure S3.