siRNA knockdown of uPAR in human bronchial epithelial cells. HBEC were treated with siRNA (negative control, uPAR specific (S032) or pooled siRNA panel (mix)) for 24 hours before RNA and protein extraction. Western blotting was performed using two different anti-uPAR antibodies and an anti-β-actin antibody. Recombinant uPAR (ruPAR) was run as a control (A). Real-time PCR was performed using a total uPAR assay and normalised using HPRT as a housekeeping gene (B). Densitometry was performed on Western blots (regions measured shown boxed) to allow semi-quantitative measurement of knockdown. Data are shown before (C) and after (D) normalisation for β-actin expression. The anti-D2 antibody detected multiple different weight species. Densitometry was performed for individual bands and normalised for β-actin expression (D).
Stewart and Sayers BMC Molecular Biology 2009 10:75 doi:10.1186/1471-2199-10-75