Anti tata binding protein

Anti-TATA-binding protein (TBP) and anti-phospho Nrf2 antibodies were purchased from Abcam, Inc. (Cambridge, MA, USA). Anti-Nrf2, anti-Akt, and anti-phospho Akt antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Fucoxanthin, anti-GCLC and anti-GSS antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium] bromide (MTT) and an anti-β-actin antibody were purchased from Sigma-Aldrich Chemical Company (St. Louis, MO, USA). Cell Tracker™ Blue CMAC was purchased from Molecular Probes (Eugene, OR, USA). LY294002 was provided by Calbiochem (San Diego, CA, USA). All other chemicals and reagents were of analytical grade.

The analysis of protein expression from in vitro B16 clones was performed as previously described [18 (link), 20 (link)–23 (link)]. The rabbit polyclonal anti myc-tag (Cell Signaling Technology, Danvers, MA, USA) and the mouse monoclonal anti-TATA binding protein (TBP) (Abcam, Cambridge, UK) primary antibodies were used in combination with the appropriate secondary antibodies HRP-conjugated (Bio-Rad). Proteins were visualised using Bio-Rad Clarity Western ECL substrate with a Bio-Rad ChemiDoc MP imaging system. Bands were quantified for densitometry using the Bio-Rad Image Lab software.

Western blot analyses were conducted according to standard protocols. The following antibodies were used: anti-FAS, anti-Nrf2, anti-NQO1, anti-Srxn1, anti-PERK, anti-JNK, anti-TGFβ1, anti-Col1a1, anti-fibronectin, anti-α-SMA, and anti-PAI1 from Santa Cruz, (Santa Cruz, CA, USA); anti-eIF2α, anti-phospho-eIF2α (Ser51), anti-CHOP, anti-ERK, anti-phospho-ERK (Tyr204/Tyr187), anti-phospho-JNK (Thr183/Tyr185), anti-p38, anti-phospho-p38 (Thr180/Tyr182), anti-Smad3, and anti-phospho-Smad3 (Ser423/425) from Cell Signaling Technology (Beverly, MA, USA); anti-phospho-PERK (Thr980) from Thermo Fisher Scientific; anti-phospho-Smad3 (Thr179) and anti-phospho-Smad3 (Ser208) from Invitrogen (Carlsbad, CA, USA); anti-TATA-binding protein (TBP) from Abcam (Cambridge, MA, USA); and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Merck Millipore. TBP and GAPDH were used as a loading control.