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4,665 Followers, 568 Following, 54 Posts – See Instagram photos and videos from p a ō l a ♡ (@paoecs) Briefly, paoecs were seeded on WillCo-dish ® glass bottom dishes (Willco Wells, Amsterdam, NL). Wild-type (siRNA-NT) and siRNA-vWF transfected cells were treated for 24 h with 100 nM AngII or saline solution (Vehicle) and then stimulated with bradykinin, as abovementioned. Briefly, paoecs were seeded at the same concentration (10 4 per well) in triplicate in 24 well plates for 6 h prior to the addition of an equal volume (50 μl) of the NADP/NADPH-Glo TM detection PAOECs transfected with siRNA-vWF. vWF gene expression was markedly reduced in transfected PAOECs and the expression of the corresponding reference gene was revealed (Suppl. File: Fig. 2). At 72h of normoxic co-culture, the level of HIF1α-exo wa s similar to single-pAOECs culture and increased by 175±11% compared to baseline (P<0.001), even if HIF1-α level in pAOECs was reduced by 40±7% than pMSCs (p<0.05). The number of both cell lines was significantly higher in co-culture compared to each monocolture. In additional experiments, the inhibition of eNOS activity by PMA did not reverse the downregulation of ET-1.ud Conclusions: We demonstrated that vWF-knockdown modulates the response of PAOECs to chronic exposure to AngII by preventing cell death, reducing ET-1 and O2- production without affecting endothelial function. . .

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