Lowering Caveolin-1 Expression in Human Vascular Endothelial Cells Inhibits Signal Transduction in Response to Shear Stress
Figure 2
Downregulation of caveolin-1 expression by RNAi.
HUVECs were treated with caveolin-1 siRNA, negative control siRNA, or left
untreated. Caveolin-1 levels were determined 48 hours after transfection. (a)
Immunoblot analysis of cell lysates. Equal loading of proteins of each
transfection condition was ensured by detecting the unrelated metabolic enzyme
GAPDH on the same blot. (b) Quantification of the signal intensities in the
immunoblot assay. The bars represent average intensities of the caveolin-1 band
in seven separate experiments. Error bars represent standard deviation.
Caveolin-1 siRNA treated cells had significantly lower caveolin-1 expression
than control siRNA-treated cells (, Student’s -test). No significant differences were found between untreated cells and negative control siRNA-treated cells. Bars represent averages of seven separate
experiments. (c) Confocal microscopic imaging of caveolin-1. Treated cells were
fixed and labeled with caveolin-1 antibodies that were detected by Alexa 633-coupled
secondary antibodies. Nuclei were stained with DAPI. All images were taken with
the same microscope settings. Scale bar is 50 m.
