Anti trx1

Cells were lysed in NETN buffer (50 mM Tris-HCl, pH 7.5; 150 mM NaCl; 1 mM EDTA; 1% NP40, phosphatase and protease inhibitor cocktail). The following antibodies were used: anti-DPF2 (Abcam, ab134942); anti-BRG1/SMARCA4 (Abcam, ab110641); anti-BAF155/SMARCC1 (Cell Signaling Technology, 11956); anti-BAF47/SNF5/SMARCB1 (Diagenode, C15410317); anti-H3 (Abcam, ab10799); anti-GAPDH (MilliporeSigma, G8795); anti-NRF2 (R&D Systems, MAB3925); anti-TRX1 (Cell Signaling Technology, 15140S); and anti-NQO1 (Cell Signaling Technology, 62262S).

The DG was extracted from the region between -1.55 mm and -2.79 mm posterior to bregma. The dissected tissue samples that were isolated using tissue punches (0.75 mm diameter) were pooled (2 to 3 samples). Subcellular fraction and western blot were described in a previous publication by our group^{26 (link)}. Anti-p-ERK1/2 (1:1,000) and anti-TRX-1 (1:1,000) were purchased from Cell Signaling Tech. Inc. (Danvers, MA, USA). β1-AR (1:500) and β2-AR (1:500) were purchased from Abcam (Cambridge, UK). Anti-class III beta-tubulin (anti-Tju1; 1:500), anti-β-catenin (1:500), anti-β-actin, and anti-lamin B were obtained from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA).

For immunoblot analyses of purified proteins or cell lysates, proteins were separated on SDS-PAGE (with or without DTT as indicated in the figures) and subsequently blotted to nitrocellulose membrane (Invitrogen) with Bio-Rad Trans-Blot® Turbo™ (25 V, 7 min). Equal loading of cell lysate proteins (30 μg per lane) was confirmed using Ponceau staining (Sigma), while chromatography fractions were instead loaded with equal volumes in each lane. Blocking was done with 5% powdered milk (Carl Roth) and 0.5% BSA (Bovine Serum Albumin, Sigma-Aldrich) in TBS-T (Tris Buffered Saline (Bio-Rad) with 0.5% Tween 20, Sigma-Aldrich). Primary antibodies were obtained from either Santa Cruz (SC) or Cell Signaling Technology (CST): anti-Trx1 (SC catalog number: 166393; 1:500), anti-TXNL1 (SC: 515218; 1:1000), anti-TrxR1 (CST#6925, 1:1000), anti-HSP27 (CST#2404, 1:1000), anti-insulin B chain (SC: 377071; 1:1000) and anti-β-actin (SC: 47778, 1:1000). The membranes were incubated overnight with the primary antibodies (diluted in TBS-T), then after washing, the corresponding HRP-conjugated secondary antibodies (Dako, P-0447, 1:8000 or Dako, P-0399, 1:3000) were added to the membranes for 1 h at RT (diluted with 5% powdered milk and 0.5% BSA in TBS-T). After addition of ECL substrate (BR1705062), chemiluminescent signals were detected using a Bio-Rad ChemiDoc XRS scanner.