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. 2006 Apr 11;94(9):1300–1310. doi: 10.1038/sj.bjc.6603094

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Copyright © 2006 Cancer Research UK

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COX-2 specific knockdown by siRNAs in HUVE cells and evaluation of 6-keto-PGF1α production. HUVE cells were transiently transfected with siRNAs directed against COX-2 mRNA (sequences A–D; final concentration 200 pM): COX-2 levels (dark bars) and 6-keto-PGF1α production (bright bars) were analysed by Western blot and ELISA assay, respectively. All procedures are described under Materials and Methods. (A) Shows COX-2 and COX-1 expression after siRNAs treatment. After the evaluation of the bands intensity by Image Master VDS software, both COX-2 and COX-1 levels were normalised against β-actin expression. All samples (lanes 2–7) except control in lane 1 were treated with PMA 40 nM. Lanes 3–6: samples treated with siRNA A, B, C and D, respectively. Lane 7: HUVE cells transfected with siRNA-Scr (scrambled), representing a negative control. Data are expressed as % of PMA-stimulated control value (lane 2) and represent the mean±s.e.m. of three independent experiments. ^*(P<0.01); #(P<0.05).