Carnosine

We used a human colon cancer cell line, Caco-2, and a human neuroblastoma cell line, SH-SY5Y. Both cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Nissui, Tokyo, Japan) supplemented with 10% fetal bovine serum (FBS; Life Technologies, Gaithersburg, MD, USA) at 37°C in 5% CO₂. Primary mouse cortex neurons (Thermo Fisher Scientific K.K., Tokyo, Japan) were cultured in complete neurobasal medium supplemented with GlutaMax-I and B-27 (Thermo Fisher Scientific K.K.) according to the instruction. Carnosine was purchased from Wako (Osaka, Japan).

An antibody against actin (SC-47778) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against CHOP (#5554) and goat anti-rabbit IgG (horseradish peroxidase- (HRP-) conjugated, #7074) were purchased from Cell Signaling Technology Japan (Tokyo, Japan). Tauroursodeoxycholic acid (TUDCA) was purchased from Tokyo Chemical Industry (Tokyo, Japan). The Zinc Assay Kit was purchased from Metallogenics Co. Ltd. (Chiba, Japan). HRP-conjugated donkey anti-mouse IgG was purchased from GE Healthcare Japan (Tokyo, Japan). Carnosine, ZnCl₂, NiCl₂, and RIPA buffer (20 mmol/L Tris-HCl (pH 7.4), 0.05 w/v% NP-40 substitute, 2.5 mmol/L MgCl₂, and 200 mmol/L NaCl) were purchased from Wako Pure Chemicals (Tokyo, Japan). Protease and phosphatase inhibitors (87786 and 78420), NuPAGE® Novex 4–12% Bis-Tris Protein Gel, iBlot™ Transfer Stack, and SuperSignal™ West Dura Extended Duration Substrate were from Thermo Fisher Scientific K.K. (Tokyo, Japan).

Murine skeletal myoblasts C2C12 (Riken Bioresource Center, Tsukuba, Japan) and human neuronal cells SH-SY5Y (ATCC, Manassas, VA, USA) were cultured in DMEM (Nissui, Tokyo, Japan) containing 10% heat-inactivated fetal bovine serum (FBS, Capricorn Scientific GmbH, Ebsdorfergrund, Germany) at 37 °C in 5% CO₂. Carnosine, AICAR and retinoic acid (RA) were purchased from FUJIFILM Wako Pure Chemical Co. (Osaka, Japan).