Figure 5.

Expression of a Zeb2 NAT is up-regulated in Snail1-transfected cells. (A) The diagram shows the sequence of NAT and the correspondence with sequences of the Zeb2 intron (shaded in gray) and the exon in 5′-UTR. The sequence within the intron corresponding to the IRES (+2357/2928) is shaded in dark gray. (B) RNA was purified from different cell lines stably transfected with control plasmids or Snail1-HA cDNA and analyzed by RT–PCR using oligonucleotides and conditions specific to Zeb2 5′-UTR NAT (see Materials and Methods). (C) The indicated fragments of the Zeb2 gene were inserted in the opposite orientation in pGL3 plasmid. The promoter activity of these DNA fragments was analyzed in RWP1 control or expressing Snail1. Values refer to that obtained with the longest fragment in RWP-1 Snail1 cells. The figure shows the average ± SD of three experiments performed in duplicate.