E cadherin sirna

SNX27-KD and WT MDA-MB-231 cells were plated in 12-well plates for 24 h before transfection. Cells were transfected with 25 nM E-cadherin siRNA (Santa Cruz Biotechnology), and control siRNA (Santa Cruz Biotechnology) using Lipofectamine-3000 transfection reagent (Invitrogen) according to the manufacturer’s instruction. The cells were harvested at 72 h post-transfection, and assayed by western blot.

3-DZNeP was purchased from Selleckchem (Houston, TX, USA). Cisplatin was purchased from Sigma (St. Louis, MO, USA). Antibodies to caspase3, E-cadherin, EZH2, H3K27me3, p-JNK, p-ERK1/2, p-p38, histone H3, and tubulin were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibody to NGAL was purchased from R&D systems (Minneapolis, MN, USA). Antibodies to GAPDH, β-actin, horseradish peroxidase-conjugated secondary antibodies, enhanced chemiluminescence (ECL) detection kit, cell counting kit-8 (CCK-8), and DAPI were from Beyotime institute of Biotechnology (Jiangsu, China). Scr and BUN reagent kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). EZH2 siRNA, E-cadherin siRNA and scramble siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Purified non-labeled mouse/rabbit mono-/polyclonal antibodies were anti-CTNNAL1, Mcl-1, CBP, p38 and p-p38 (Abcam, Cambridge, MA, USA), ERK, p-ERK, JNK, p-JNK, c-Jun, p-cJun, p21^waf/cip1, p53, GAPDH, (Cell Signaling Technology, Inc., Danvers, MA, USA), KI-67 (Santa Cruz, Texas, USA). HRP-conjugated secondary anti-mouse and anti-rabbit antibodies were obtained from Life Technologies and Cell Signaling Technology, Inc., respectively. Anti-α-tubulin-HRP conjugated antibody was purchased from Cell Signaling Technology, Inc.
Texas-Red conjugated secondary anti-mouse and anti-rabbit antibodies were purchased by Jackson Immuno (Newmarket, UK). HGF and TNF-α were purchased from PeproTech Inc. (Rocky Hill, NJ). LPS was purchased from Sigma-Aldrich (St. Louis, MO, USA).
5x NFκB-luc-Reporter and AP-1-luc-Reporter plasmids were obtained from Agilent technologies (Santa Clara, USA), pGL3 reporter and control vectors were purchased from Invitrogen, E-cadherin si-RNA was purchased from Santa Cruz Biotechnology, Inc. and Caspase3/7, 8 and 9 assay and CellTiter-Blue cell viability Assay from Promega (Madison, USA). Staurosporine was purchased from eBioscience (CA, USA). Chemotherapeutic agents dacarbazine, cisplatin and paclitaxel were acquired from Sigma-Aldrich.