Anti tnfr2

The protein expression levels were evaluated by Western blotting. Briefly, 1 × 10⁶ cells in 60 mm culture dish (BD Falcon) were incubated with the indicated concentrations of SAHA for 24 hours. Then cells were washed with PBS and added in 4 volumes of lysis buffer (Intron Biotechnology, Seongnam, Gyeonggi-do, Korea). Thirty μg of total protein were resolved by 4–20% SDS-PAGE gels, and then transferred to Immobilon-P PVDF membranes (Millipore) by electroblotting. Then membranes were probed with anti-PARP, anti-c-PARP, anti-Bax, anti-Bcl-2, anti-TNFR2 (Cell signaling Technology, Danvers, MA), anti-TNFR1, anti-β-actin and anti-GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA). Membranes were incubated with horseradish peroxidase-conjugated secondary antibodies. Blots were developed using an EZ-Western Lumi Pico ECL solution kit (DoGen, Seoul, Korea).

Staurosporine (STS), an apoptosis inducer, and U0126, an inhibitor of MEK, were purchased from Enzo Life Sciences, Inc. (Farmingdale, NY, USA). Etoposide (ETO), an apoptosis inducer, was purchased from Tokyo Chemical Industry Co., LTD. (Tokyo, Japan).
The following antibodies were used for western blot, immunoprecipitation, and immunofluorescence (IF) analysis: anti-N-cadherin (BD Transduction Laboratories, Franklin Lakes, NJ, USA), anti-Flag (Sigma Aldrich, MO, USA), anti-DR-5, anti-DcR-2, anti-Fas, anti-TNFR-1, and anti-TNFR-2 (Cell Signaling Technology, Inc., Danvers, MA, USA), anti-phospho extracellular signal-regulated kinase (ERK)1/2, anti-ERK1/2 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-phospho p38, anti-p38, anti-phospho Akt, anti-Akt, anti-phospho NF-kB p65, anti-NF-kB p65, anti-cleavage PARP (Cell Signaling Technology, Inc.), and anti-β actin (Sigma-Aldrich Corporation, St. Louis, MO, USA).
HOC313 and HOC719NE cells were transfected with N-cadherin small interfering RNA (si-CDH2) (Ambion, Life Technologies, Foster City, CA, USA) using Oligofectamine reagent (Invitrogen Corporation) following the manufacturer’s instructions.