Fig. 6.

SIA cleavage attenuates eNOS phosphorylation by USS and induces VCAM-1. HUVEC were incubated with neuraminidase (Neur., 2 U ml^-1, 30 min) before exposure to USS (15 dyn cm^-2) for the indicated time points. (A-B) Representative immunoblots and densitometric analyses of eNOS phosphorylation at S1177 and S633 relative to β-actin and total eNOS levels. Data denote mean ± S.E.M. (n = 4–6 different donors). *P < 0.05 (2-way ANOVA). (C) Representative immunoblot and densitometric analysis of VCAM1 expression relative to β-actin. Data denote mean ± S.E.M. (n = 3 donors). *P < 0.05, **P < 0.01 (2-way ANOVA). (D) Representative images of p65 subunit cellular distribution (green) and quantification of nuclear-to-cytosolic fluorescence intensity in fixed cells. Cell nuclei were co-stained with DAPI which was omitted for clarity. Data denote mean ± S.E.M. (n = 4 different donors) of fluorescence intensity values from at least 300 cells per condition. *P < 0.05 (2-way ANOVA). Scale bar=20 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)