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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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Small interfering RNA-B (siRNA-B) activates the interferon-signalling cascade in HUVEC only at high concentration. Cells were transiently transfected with siRNA-B directed against COX-2 mRNA as described under Material and Methods. Two different final concentrations were used (200 nM and 200 pM). Phospho-STAT-1 (Tyr701) and STAT-1 proteins expression was analysed by Western blot (A) and pSTAT-1 levels, normalised with respect to STAT-1 total levels, are reported in (B). Samples in lanes 2, 4 and 6 were treated with PMA 40 nM and represent positive controls. Lanes 1: negative control (no PMA stimulation). Lanes 3 and 4: samples treated with siRNA-B 200 nM. Lanes 5 and 6: samples treated with siRNA-B 200 pM. Data are expressed as % of positive control value in lane 2 and represent the mean±s.e.m. of three independent experiments. The same treatments were used in an immunofluorescence assay to determine the phospho-STAT-1 protein levels and localisation in siRNA-transfected HUVE cells (results are shown in C). ^*(P<0.01). Bar: 20 μm.