Anti grhl2

We performed western blotting as described previously [39 (link)]. The following primary antibodies were used for protein detection: anti-TET2 (catalog number 61390, Active Motif, Carlsbad, CA), anti-ESRP1 (catalog number GTX131373, GeneTex, Irvine, CA, USA), anti-CD44 (catalog number MAB7045, R&D Systems, Minneapolis, MN, USA), anti-ADARB2 (catalog number sc-73410, Santa Cruz Biotechnology, Dallas, TX, USA), anti-ZEB1 (catalog number 3396, Cell Signaling Technology, Danvers, MA, USA), anti-GRHL2 (catalog number HPA004820, Sigma-Aldrich, St. Louis, MO), and anti-vimentin (catalog number 3932, Cell Signaling Technology). The ECL prime blocking agent (GE Healthcare Life Sciences, Piscataway, NJ, USA) was used for blocking and Lumigen TMA-6 reagents for detection (Lumigen, Inc., Southfield, MI, USA). The blots were re-probed with an HSP90 antibody (catalog number 4875, Cell Signaling Technology) or with an anti-β-actin antibody (catalog number A5316, Sigma-Aldrich) to document equal protein loading. Each western blot was performed at least twice; representative blots are shown. Relative intensities of the bands were determined with ImageJ software (National Institutes of Health, Bethesda, MD, USA).

We separated proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and detected various proteins by Western blotting, as described previously [42] (link). The following primary antibodies were used for detection: anti-COX-2 monoclonal antibody (Cayman Chemical, Ann Arbor, MI), anti-GRHL2 (Sigma-Aldrich, St. Louis, MO), anti-FTO (EMD Millipore, Billerica, MA), anti-ZEB1, anti-histone H3, anti-histone H3 tri-methyl Lys-4, anti-histone H3 acetyl Lys-14, and anti-vimentin antibodies (Cell Signaling, Danvers, MA). We used the ECL prime blocking agent (GE Healthcare Life Sciences, Piscataway, NJ) for blocking and Lumigen TMA-6 reagents for detection (Lumigen, Inc., Southfield, MI). The filters were stripped by incubating the membrane in 0.5% Triton X-100 and were re-probed with a monoclonal β-actin antibody (Sigma-Aldrich, St. Louis, MO) or with vinculin antibody (Abcam, Cambridge, MA), which served as gel-loading controls. To detect β-actin or vinculin, we used 2% non-fat dry milk (Bio-Rad, Hercules, CA) for blocking and ECL prime reagent for detection (GE Healthcare Life Sciences, Piscataway, NJ). We performed each western blot at least twice; the representative blots are shown. We quantified the protein bands on x-ray films by using the ImageJ image processing program (National Institutes of Health).