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Dulbecco’s modified Eagle’s medium (DMEM), penicilin-streptomycin, and trypsin-ethylenediaminetetra acetic acid (EDTA) were purchased from GIBCO-BRL (Grand Island, NY, USA). Fetal bovine serum (FBS) was purchased from Hyclone (Logan, UT, USA). Gelatin, PI, and MTT were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies specific to caspase-3, caspase-8, caspase-9, cFILP, cIAP-2, Bcl-XL, COX-2, phospho-NF-κB, phospho-c-Jun, c-Jun, phospho-IKK, phospho-IκB, phospho-p38, p38, phospho-JNK, JNK, phospho-ERK1/2, ERK1/2, phospho-AKT, AKT, phospho-TAK, TAK, and RIP were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). NF-κB, PARP, MT1-MMP, uPAR, ICAM-1, and VEGF were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Cyclin D1 was purchased from Milipore. The Can Get Signal^® Immunoreaction Enhancer Solution was purchased from Toyobo (Osaka, Japan). Matrigel was purchased from Becton Dickinson (Bedford, MA, USA). Dicentrine was ordered from Chengdu Biopurify Phytochemicals Ltd. (Sichuan, China).

Prostate CAFs were seeded in 6-well plates at a density of 3 × 10⁵ cells/well and were treated with MPSSS at different concentrations (0, 0.1, 0.2, 0.3, 0.4, and 0.5 mg/ml) for 24 h. Then the cells were lysed with RIPA buffer supplemented with 100 mΜ phenylmethylsulfonyl fluoride (PMSF), 25 μg/ml aprotinin, 1 mM sodium orthovanadate, and 50 nM NaF to obtain the total protein content. The total protein concentrations were determined by the BCA protein assay. Equal amounts of protein samples (30 μg/sample) were separated on 10% SDS polyacrylamide gels under denaturing conditions and were then electrotransferred onto nitrocellulose membranes (GE Healthcare, Milwaukee, WI, U.S.A.) for 70 min at 100 V. Then, the membranes were blocked with 3% BSA in PBS-T (0.1% Tween-20) for 1 h and were incubated overnight at 4°C with the primary antibodies. The primary antibodies used were against α-SMA, phospho-NF-ĸB p65, NF-ĸB p65, phospho-TAK, TAK, phospho-IKKα/β, TRAF6, and MyD88 (all from Cell Signaling Technology, U.S.A.) and were diluted at 1:1000; the primary antibody against GAPDH (Tianjin Sungene Biotech Co., Ltd.) was diluted at 1:2000 and was used as an internal standard.