Tead4 ab58310

Extracted samples were separated by 12% SDS-polyacrylamide gel electrophoresis (PAGE) (Bio-Rad) and transferred to nitrocellulose membranes. Antibodies for western blot analysis were as follows: mouse monoclonal anti-Myc (9E10) from Roche (Basel, Switzerland); mouse monoclonal anti-Flag (F1804) and mouse monoclonal anti-β-actin (AC-15) from Sigma; mouse monoclonal YAP antibody (63.7) from Santa Cruz Biotechnology (Dallas, TX, USA) and mouse monoclonal TEAD4 (ab58310) from Abcam (Cambridge, United Kingdom). First antibody incubation was followed by probing with the corresponding secondary antibody and developing using Amersham Imager (Little Chalfont, United Kingdom). Uncropped images of western blottings are available in Figure S3.

Western blotting was performed using an enhanced chemiluminescence kit (Merck Millipore), as described previously.¹⁵ The following antibodies were used: myc (PL14) and β‐actin (M177‐3) from MBL (Nagoya, Japan); E‐cadherin (#3195), N‐cadherin (#13116), Smad2/3 (#8685), phospho‐Smad2 (#3108), phospho‐Smad3 (#9520), Snail (#3879), Slug (#9585), HMGA2 (#5269) from Cell Signaling Technology (Beverly, MA); TEAD4 (ab58310) from Abcam (Cambridge, UK); GFP (mFX75) from Fujifilm Wako (Tokyo, Japan); TetR (TET01) from MoBiTech (Cambridge, MA, USA); and TWIST (25465–1‐AP) from Proteintech (Rosemount, IL, USA). Horseradish peroxidase (HRP)‐F(ab’)2 secondary antibodies were purchased from GE Healthcare (Waukesha, WI, USA). Protein bands were analysed using a ChemiDoc XRS+image analyzer (Bio‐Rad, Hercules, CA, USA). The intensity of bands was measured by Quantity One software (Bio‐Rad). Quantitative ratios of E‐cadherin, N‐cadherin, Snail, Slug, HMGA2 or TEAD4 to actin and EGFP to TetR, which were calculated based on the data, are shown as relative values.