Fig. 3.

RNAi-mediated knockdown of PEA3 results in upregulation of Fgf-10 expression. Transfection with RNAi targeted to the PEA3-coding sequence resulted in a 64% drop in Pea3 expression levels, by 24 h and persisting at 48 h, in immortalised mouse lung endothelial cells (A). Cells transfected with a scrambled RNAi control pool showed a level of Pea3 expression identical to that seen in mock-transfected cells (data not shown). This decrease in PEA3 mRNA level was mirrored by a decrease of 34% in the level of PEA3 protein in cell lysates prepared 48 h post-transfection (B). By 48 h post-transfection, cells treated with PEA3 RNAi showed a 2.6-fold increase in Fgf-10 expression, as measured by real-time RT-PCR and analysed using the 2^−ΔΔCt method (C). All quantitation represents data from at least three independent experiments, and reflects either real-time RT-PCR data (n⩾3; each in triplicate) (A, C) or Western blot densitometry (n=3) (B). Statistical significance was determined with Student's t-test or Mann-Whitney Rank Sum test (PEA3 mRNA expression after 48 h). Results were considered significant at P<0.05 (*).