Figure 3.

The increased VEGF secretion in DKC1 KD cells is due to the up-regulation of VEGF mRNA IRES-mediated translation. (A) Histograms show the level of VEGF protein detected using an ELISA array. Experiments were performed in MCF-7 control (left) and DKC1 KD (right) cells treated with 15 nM PTC299 for 72 h. VEGF secretion of PTC299 treated cells is normalized on the correspondent untreated cells; for the effect of DKC1 KD on VEGF secretion on MCF7 cells see Figure 2A (B) VEGF-IRES–mediated translation measured in MCF7 control and DKC1 KD cells after 72 h of 100 nM PTC299 treatment. siRNA transfection was performed 96 h before cell harvesting. (C) Representative image (left) and a summarizing graph (right) from clonogenic assays performed with control (empty vector) and shDKC1 MCF7 cells. Histograms represent means and SDs from three independent experiments. P < 0.05 is considered significant. NS = not significant.