Fig. 1

RT-PCR and western blotting results validated the efficacy of shRNAs to silence the expression of HMGA2 gene in ACHN cells. A RT-PCR analysis of the HMGA2 mRNA expression in ACHN cells receiving either a scrambled control shRNA or a shRNA (#1-#3) specific to HMGA2 gene (statistical significance with p = 0.002, p < 0.001, p = 0.002 for the three shRNA 1–3). B Western blotting analysis of the HMGA2 and GAPDH protein expression in ACHN cells receiving HMGA2 shRNA2. C Quantification was performed by normalizing the protein expression level of HMGA2 against the level of the house-keeping gene GAPDH (p = 0.002**). D the mRNA expression level of HMGA2 in stable-transfected and puromycin-selected ACHN cells (p = 0.013*). This is done before xenograft modeling. E-G The mRNA expression level of EMT markers E- cadherin, N-cadherin and Snail in cultured ACHN cells (p = 0.0132 *, p = 0.0102 *, p = 0.0024 **, for the three groups). H Western blot results for protein expression of EMT-related genes in HGMA2-silenced ACHN cells before xenografting. Data were expressed as mean +/− standard deviation. For each group, data were obtained from three independent repeat experiments, with triplicate samples in each repeat. Statistical significance was calculated using Kruskal-Wallis test