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. 2006 Apr 25;94(10):1516–1523. doi: 10.1038/sj.bjc.6603108

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Copyright © 2006 Cancer Research UK

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Silencing of PAIP2 by RNA interference inhibits VEGF-A expression in a human pharynx carcinoma cell line. (A) Total protein extracts from Detroit 562 cells transiently transfected twice with siRNAs against PAIP2 or GFP, were analysed by Western blotting using a polyclonal anti-PAIP2 antibody. The amounts of ERK2 are shown as a loading control. (B) Total RNA extracted from the same cells was subjected to Northern blotting analysis for VEGF-A mRNA expression (upper panel). The arrows point to the different VEGF-A isoforms. 36B4 RNA is shown as a loading control. The levels of VEGF-A mRNA were quantified by phosphorImaging analysis (lower panel). The values are normalised to 36B4 RNA. Values obtained with the control siRNA are taken as 100%. The data shown represent the means and standard errors of two independent experiments. (C) Tissue culture supernatants from the same cells were analysed using a human VEGF-A specific ELISA. The values are normalised to the cell number. Values obtained with the control siRNA are taken as 100%. The data shown represent the means and standard errors of two independent experiments each performed in triplicate.