Ctbp1

The Western blotting was performed as previous studies.²³, ²⁴ Briefly, cell samples were mixed with lysis buffer (0.01 M Tris‐HCl, pH 7.4; KCl, 0.15M; NaF, 0.1M; EDTA, 0.002M; β‐mercaptoethanol, 0.012 M; Nonidet P‐40, 0.5%) and lysed for 2 hours at 4℃ in the presence of a mixture of protease inhibitors (leupeptin, 0.01 mg/mL; Na₃VO_4, 0.001M; Pefabloc, 0.3 mg/mL; okadaic acid, 0.01 μM). The proteins were mixed with sample buffer to denature and heated at 95°C for 1 min, separated through SDS‐PAGE (10% polyacrylamide gel) and transferred onto membranes of PVDF. Membranes were blocked with fat‐free milk (5% in TBST: 50 mM Tris/pH 7.5, containing 0.15M NaCl and 0.05% Tween‐20) for 60 min at room temperature and probed overnight with primary antibodies at 4°C. The blots were washed, kept in the secondary antibody for 60 min at room temperature, the bands observed and evaluated through UVP Bio Imaging systems. The primary antibodies used are as list: CtBP1 (ab129181, Abcam, Cambridge, MA, USA), N‐cadherin (ab76011, Abcam), E‐cadherin (ab15148, Abcam), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), CCL2 (@2027, Cell Signaling Technology), p65 (#8242, Cell Signaling Technology), p‐p65 (#3033, Cell Signaling Technology), Lamin A/C (#4777, Cell Signaling Technology) and β‐actin (A5441, Sigma, St. Louis, MO, USA).