Phospho akt 4060p

SFA gels was provided by Guiyang Xin Tian Pharmaceutical Co., Ltd. (lot number JG141002; Guizhou, China). Matrine (Lot NoMKBS8661V) and Oxymatrine (Lot No 011M1378V) were purchased from Sigma. Antibodies against AKT (4691, 1:1000), Phospho-AKT (4060P, 1:1000), E-Cadherin (3195,1:1000), MMP2 (13132, 1:1000) and mTOR Substrates Sampler kit (CST 9862), which includes Phospho-mTOR, mTOR, were purchased from Cell Signaling Technology. Antibodies against Bcl-2 (ab32124, 1:1000), Bax (ab32503, 1:1000), Cyclin A (ab181591, 1:2000), CDK2 (ab32147, 1:1000) were from Abcam.

Gemcitabine was purchased from Lilly France (St-Cloud, France) (Lot No A875303A). Antibodies against β-catenin (51067-2-AP, 1:800), LC3A/B (66139-1-1g, 1:1,000), Cyclin D1 (60186-1-1g, 1:1,000) and GAPDH (60004-1-Ig, 1:1,000) were from Proteintech Group, Inc., (Chicago, IL, USA). Antibodies against MEK1/2 (9126, 1:1,000), Phospho-MEK1/2 (2338, 1:1,000), Phospho-ERK1/2 (4376, 1:1,000), AKT (4691, 1:1,000), Phospho-AKT (4060P, 1:1,000), Phospho-P38 (4511, 1:1,000), Phospho-JNK (4668, 1:1,000), E-Cadherin (3195, 1:1,000), Phospho-p53 (9284, 1:1,000), and mTOR Substrates Sampler kit (CST 9862), which includes Phospho-mTOR, mTOR, Phospho-p70s6389 (Thr389), Phospho-P70389 (Thr371), Phospho-4EBP1 (Thr37/46), and anti-rabbit IgG, were purchased from Cell Signaling Technology, Inc., (Danvers, MA, USA). Antibodies against p38 (sc-7149, 1:1,000), P53 (sc-126, 1:1,000) and JNK (sc-571, 1:1,000) were from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). PD98059 (MEK inhibitor, 9900, 20 UM), rapamycin (mTOR inhibitor, 9904, 10 nM), and LY294002 (PI3K Inhibitor, 9901, 50 μM) were purchased from Cell Signaling Technology.