Anti trx

Bicinchoninic acid (BCA), trichloroacetic acid (TCA), sulforhodamine B (SRB), propidium iodide, RNase A, dimethyl sulfoxide (DMSO) and catalase were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). Roswell Park Memorial Institute (RPMI) 1640 medium, Dulbecco Modified Eagle Medium (DMEM)/high glucose, Minimum Essential Media (MEM), fetal bovine serum (FBS), trypsin-EDTA solution (1X), antibiotic-antimycotic solution (100X), and PBS (1X) were purchased from HyClone Laboratories, Inc. (South Logan, UT, USA). Anti-caspase-9, anti-cleaved caspase-9, anti-caspase-3, anti-BID, anti-p-p53 (Ser15), anti-Prx1, anti-caspase-8, and anti-cleaved caspase-8 were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-cleaved caspase-3, anti-Bax, anti-Bcl-2, anti-p53, anti-catalase, anti-SOD-1, anti-SOD-2, anti-GPx-1, anti-Trx, anti-PrxI/II, anti-β-actin, and all secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-BIM, anti-PARP, anti-cleaved PARP, and fluorescein isothiocyanate (FITC) Annexin V apoptosis detection kit I were purchased from BD Biosciences (CA, USA). Anti-RIP-3 was purchased from Abcam (Cambridge, MA, USA). 2′,7′-dichlorodihydrofluorescein diacetate (DCFH₂-DA) was purchased from Invitrogen (Carlsbad, CA, USA).

Immunoblot analyses were carried out as described previously (13 (link)). Digital images were captured using LAS-1000 Plus (Fujifilm). The following primary antibodies were used: monoclonal anti–Myc 9E10 (Santa Cruz Biotechnology, sc-40), anti–p38α F9 (sc-271120), anti–JNK2 D-2 (sc-7345), anti–ASK1 F-9 (sc-5294); anti–β-actin 6D1 (Wako, 010-27841); anti-Flag M2 (Sigma-Aldrich, F1804); anti–phospho-p38 D3F9 (Cell Signaling Technology, 4511), anti–phospho-JNK 81E11 (4668), anti–phospho-JNK 98F2 (4671), anti–phospho-MAPKAPK2 27B7 (3007), anti–phospho–c-Jun 54B3 (2361); polyclonal anti-Flag (Sigma-Aldrich, F7425); anti-JNK (Cell Signaling Technology, 9252), anti–phospho-MKK3/6 (9231), anti–phospho-MKK4 (9156), anti–phospho-MKK7 (4171), anti-MKK7 (4172), anti-MAPKAPK2 (3042); anti-Trx (Santa Cruz Biotechnology, sc-20146), anti-Myc (sc-789), anti-p38α (sc-535), anti-JNK (sc-571), anti-MKK3/6 (sc-13069), anti-MKK4 (sc-13070), anti-MTK1 (sc-28770), anti–c-jun (sc-1694); and anti-GADD45β (Cloud-Clone, PAL535Hu01). Affinity-purified anti-MTK1 and anti–phospho-MTK1(T1493) antibodies were made in-house (13 (link), 15 (link)). All antibodies were used at a dilution of 1:1000, except anti-ASK1 (1:500) and anti-GADD45β (1:400).