Western blot analysis and chemicals
Western blotting was performed as described previously32. Mouse tissue and cells were lysed in RIPA [50 mM Tris–HCl (pH 7.5), 150 mM NaCl, 0.1% sodium dodecyl sulfate (SDS), 0.1% sodium deoxycholate, and 1% Nonidet P-40] with Roche cOmplete™ mini tablets (Roche, Mannheim, Germany), 25 µM MG132 (EMD Chemicals, Inc; San Diego, CA), and phosphatase inhibitors, [20 mM β-glycerophosphate (Sigma–Aldrich Co., LLC; St. Louis, MO) and 20 mM sodium orthovanadate (Fujifilm, Osaka, Japan)]. After centrifugation at 13,100 × g, the supernatants were used for western blotting with the following antibodies: anti-CHOP (Thermo Fisher Scientific, Waltham, MA; Cell Signaling Technology, Inc; Danvers, MA), anti-ATF4 (Santa Cruz Biotechnology, Inc; Santa Cruz, CA; Thermo Fisher Scientific), anti-p35 (C-19), anti-GAPDH (Santa Cruz Biotechnology, Inc),…
