Chunk #3 — Materials and Methods — Reagents

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Alcohol stimulates activation of Snail, epidermal growth factor receptor signaling, and biomarkers of epithelial-mesenchymal transition in colon and breast cancer cells.

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Alcohol (ethanol) solutions were made daily (AAPER Alcohol and Chemical Co., Shelbyville, KY). TACE inhibitor: TAPI-2 (TNF-α Protease Inhibitor-2) (Chemicon, Temecula, CA). EGFR inhibitor: AG1478 (Calbiochem, San Diego, CA). All EGFR, MAPK (ERK1 and ERK2), and Histone H3 Abs were from Cell Signaling Technology (CST) (Danvers, MA). ERK1/2 (MEK1) inhibitor: PD98059 (Calbiochem). β-actin, MMP-2, MMP-7, and MMP-9 Abs: Sigma-Aldrich (St. Louis, MO) and MMP-7 positive control (Chemicon). Phospho-Snail Ser246 Ab: Abcam (Cambridge, MA), and total Snail Ab: CST. TNF-α: R&D Systems (Minneapolis, MN). Vimentin mouse Ab: Santa Cruz Biotechnology (Santa Cruz, CA) and secondary Abs were anti-mouse Alexa fluor 488 and anti-rabbit Alexa fluor 594 (Invitrogen, Carlsbad, CA).