Fig 1: Validation of candidate Snail1 target proteins.(A) Western blot analysis of the expression of selected proteins in cytosolic (C) and nuclear (N) extracts of SW480-ADH Mock and Snail1 cells. HNRNPH3 isoforms (1–4) are indicated. The numbers below the gels indicate the quantification of the change in expression between Snail1 and Mock cells after normalization to ß-Tubulin or Lamin B (cytosolic and nuclear control, respectively). The statistical significance of the nuclear expression changes promoted by Snail1 was assessed by two-tailed unpaired Student's t-test. It was p<0.001 for 14-3-3e, 14-3-3s, 14-3-3?, CPSF6 and the isoforms 3 and 4 of HNRNPH3; p<0.01 for 14-3-3t; and p<0.05 for 14-3-3ß and PA2G4. The regulation of SFPQ (p = 0.057) and that of the isoforms 1+2 of HNRNPH3 (p = 0.058) did not reached the statistical significance, but a clear tendency was observed. (B) Western blot analysis of the expression of selected proteins in cytosolic (C) and nuclear (N) fractions of HT29 Mock and Snail1 cells. The numbers below the gels indicate the quantification of the change in expression between Snail1 and Mock cells after normalization to ß-Tubulin or Lamin B (cytosolic and nuclear control, respectively). (C) Quantitative RT-PCR study of PA2G4 expression in SW480-ADH and HT29 Mock and Snail1 cells. 18S rRNA expression was used for normalization. Statistical significance was assessed by two-tailed unpaired Student's t-test, single asterisk indicates p<0.05.