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. 2014 Mar 14;9(3):e91346. doi: 10.1371/journal.pone.0091346

Figure 3. Inhibition of PI3 kinase does not affect S1P-induced WPB release.

Figure 3

(A/B) HUVECs were transfected with a control siRNA SMARTpool (siCTRL) or a siRNA SMARTpool targeting S1PR1 (siS1PR1) or S1PR3 (siS1PR3). Transfected HUVECs were then incubated for 60 minutes with SF medium or SF medium containing 10 μM LY294002. After preincubation, HUVECs were stimulated for 45 minutes with 1 μM S1P, or SF medium alone (-) in presence or absence of PI3K inhibitor LY294002. The amount of VWF secreted in the medium was measured by ELISA. Total amount of VWF secreted is displayed in percentages; the amount of VWF secreted by unstimulated cells at 45 minutes in cells transfected with siCTRL was set to 100%. Three independent experiments were performed. Statistical significance was assessed by 2-way ANOVA followed by Bonferroni post-hoc test for selected comparison (***P<0.001; **P<0.01 and *P<0.05).