Figure 1.

Zeb2 protein is increased in Snail1 transfectants by a post-transcriptional mechanism. (A) Total cell extracts were prepared from HT-29 M6, SW-480, LS-174T, or RWP-1 cells, control or transfected with Snail1-HA cDNA (see the Materials and Methods). One-hundred-fifty micrograms of protein from total cell extracts were analyzed by Western blot using a mab specific to Zeb 2 protein. mRNA was isolated from the indicated cell lines and analyzed by qRT–PCR (C) or semiquantitative RT–PCR (D), with the oligonucleotides depicted in B. Oligos 3F–2R (C) corresponded to the translated sequence, 1F1R (D, bottom) flanked the intron present in the 5′-UTR, and 2F–1R (D, top), determined the conservation of the intron. The size of the DNA fragments amplified is indicated. The figure shows the result of one experiment out of three performed. In B, the ORF is depicted in black, the 5′-UTR and 3′-UTR exons are in gray, and the 5′-UTR intron is in white.