Real Time PCR for EWS-FLI1 target genes and genes involved in EMT:A: Real time PCR for target genes showing down regulation of EWS-FLI1 regulated genes like Cyclin D1, c-MYC, GLI1, NKX2.2, EZH2, NROB1 in junction construct (aa 251–280) transfected EWS502 cell line relative to vector control pcDNA-EWS502. Figure 6B: Real Time PCR for EMT markers showing down regulation Fibronectin, Slug, Vimentin, N-Cadherin and up regulation of E-Cadherin in junction region transfected EWS502 cell line relative to vector control pCDNA-EWS502. Figure 6C: Immunoblot analysis of E-Cadherin, Vimentin and Beta Actin expression in MCF-7, pCDNA/FLAG and pCDNA/FLAG/Junction (aa 251–280) stably expressing EWS502 cells. Figure 6D: Quantitation of Immunoblot intensities. The individual intensities are presented as percent total intensity . The total intensity was obtained by adding all the intensities from the individual bands.
Jully et al. BMC Cancer 2012 12:513 doi:10.1186/1471-2407-12-513