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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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Effect of anti-COX-2 shRNA expression on the interferon-signalling cascade in HT29 pSUPER(+) cells. An analysis of STAT-1 phosphorylation in HT29 wild-type cells and HT29 cells infected using pSUPER.retro system was performed by Western blot (A and B) and immunofluorescence (C) assays, following the procedures described under Material and Methods. 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). Lanes 1 and 2: HT29 wild type. Lanes 3 and 4: HT29 pSUPER(+). Samples in lanes 2 and 4 were treated with PMA 40 nM. Data are expressed as % of positive control value in lane 2 and represent the mean±s.e.m. of three independent experiments. The same samples were analysed in an immunofluorescence assay to determine the phospho-STAT-1 protein level and localisation (results are shown in C). ^*(P<0.01). Bar: 20 μm.