Comparative expression data between wild-type and ANGPTL4-knockdown MKN28 and MKN28snai1ER cells upon EMT induction

Epithelial-mesenchymal transition (EMT) induced by microenvironment stimuli can be attributed to the transcriptional regulation of epithelial and mesenchymal phenotypes. Here we show how EMT is coordinated with cancer metabolism, an emerging hallmark of tumorigenesis. To identify molecular drivers of EMT common to all three EMT models, we performed a comparative microarray gene expression analysis. Cells were seeded at a density of 5.25×102 cell/cm2 to allow the formation of individual colonies before indicated treatments. Hypoxia treatments were performed in hypoxic chamber (Stem cell Technology, USA) purged with 5% CO2 and 95% N2 to obtain O2 concentration at 1% as determined by an O2 sensor. For TGF-β-induced EMT, cells were treated with 10 ng/mL of TGF-β for 2 days. For Snai1-mediated EMT, MKN28Snai1ER/shANGPTL4 and MKN28Snai1ER cells were exposed to 20 ng/mL of 4-hydroxytamoxifen (4-OHT) for 4 days. RNA was extracted from the respective treatments with Trizol following the manufacturer’s protocol. Further sample processing of the RNA was carried out using Applause® WT-Amp ST System (NuGEN), and microarray experiments were performed on GeneChip® Human Gene 1.0 ST arrays according to the manufacturer’s instructions.