Nuclear-Cytoplasmic Fractionation and Western Blotting

Nuclear/cytoplasmic fractionation was separated using the Cell Fractionation Kit (Cell Signaling Technology, USA) according to the manufacturer’s instructions, and the whole cell lysates were extracted with RIPA Buffer (Cell Signaling Technology). Western blotting was performed according to a standard method, as described previously [35 (link)]. Antibodies against E-cadherin (Cat# 3195), Vimentin (Cat# 5741), Fibronectin (Cat# 4706), SOCS1 (Cat# 3950), TNIP1 (Cat# 4664) and PIAS4 (Cat# 4392) were purchased from Cell Signaling Technology, and p65 (cat# 10745–1-AP) from Proteintech, p84 (Cat#:PA5–27816) from Invitrogen and PDLIM7 (Cat#:SAB1406807) from Sigma-Aldrich,USA. The membranes were stripped and reprobed with an anti–α-tubulin antibody (Sigma-Aldrich, USA) as the loading control.