Ptp1b

Protein lysates were acquired using RIPA lysis buffer supplemented with both phosphatase and protease inhibitor cocktails (Life Technologies, Thermo Fisher Scientific brand, Waltham, MA, USA). Equal amounts (5 μg) of proteins in RIPA buffer were separated on 10% NuPage Bis-Tris gels (Invitrogen) and transferred to a PVDF membrane (Invitrogen). Antibodies used are as follows: E-Cad (BD), PTP1B (BD, Fisher Scientific brand, Waltham, MA, USA), pSrc529 (Abcam), claudin-1 (Zymed, San Francisco, CA, USA), occludin (Zymed), β-catenin (Biolegend, San Diego, CA, USA), FAK (Cell Signaling, Danvers, MA, USA), pFAK (Cell Signaling) GAPDH (Abcam) and actin (Abcam) were used. In Figures 1,4 and 5, secondary antibodies were horsesadish peroxidase-conjugated anti-mouse or rabbit (BD Scientific; Fisher Scientific brand, Waltham, MA, USA) used at dilution 1:10 000. The protein bands were visualized using enhanced chemiluminescence (ECL) system (Thermo Scientific, Waltham, MA, USA). In Supplementary Figure 4, secondary antibodies were mouse or rabbit IRDye (Li-Cor, Lincoln, NE, USA) used at 1:10 000 and protein bands were detected using the Odyssey Infrared Imaging System (Li-Cor) and then fluorescent images were converted to gray scale.
All western blots are representative of three separate experiments.