Effect of shear stress on nuclear translocation of NFκB. (A) Nuclear extract of HUVECs was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to PVDF membrane by Western blotting. Then, p65 subunit of NFκB was detected with rabbit anti-p65 polyclonal antibody. (B) Cytoplasmic extract of HUVECs was separated by SDS-PAGE and transferred to PVDF membrane by Western blotting. Then, IκBα was detected with rabbit anti-IκBα polyclonal antibody. Lane 1, static control; lane 2, cells stimulated with TNF-α (100 U/mL) for 30 minutes; lane 3, cells exposed to shear stress at 18 dynes/cm2 for 15.5 hours; lane 4, cells exposed to shear stress at 18 dynes/cm2 for 15 hours and then stimulated with TNF-α (100 U/mL) and sheared for a further 30 minutes; lane 5, cells exposed to shear stress at 18 dynes/cm2 for 30 minutes.
