Fig. 4.
Endogenous NEU1 knockdown enhances Nrf2-mediated antioxidant signaling. EA. hy926 cells transduced with lentiviral particles either containing sialidase 1 (NEU1) silencing shRNA (LvNEU1) or scrambled sequences (Scr) were exposed to USS (15 dyn cm-2), OSS (±5 dyn cm-2, 1 Hz) or maintained in static culture for 48 h. (A) Representative immunoblot and densitometric analysis of NEU1 expression in whole cell lysates presented relative to β-actin. Data denote mean ± S.E.M. (n = 5 independent cultures). *P < 0.05; **P < 0.01; ***P < 0.001 (2-way ANOVA). (B) Representative fluorescence images of the SIA component of the GCX stained with WGA-CFTM448A (red) in fixed cells using an inverted epi-fluorescence microscope. WGA mean cell intensity (MCI) was normalized to the respective number of cell nuclei stained with DAPI (blue). Data are expressed as mean ± S.E.M. (n = 3 independent cultures). Scale bar=20 μm. (C) Representative immunoblots and densitometric analyses of HO-1, NQO1 and GCLM expression in whole cell lysates presented relative to β-actin. Data denote mean ± S.E.M. (n = 5 independent cultures). *P < 0.05; **P < 0.01; ***P < 0.001 (2-way ANOVA). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)