Amg9810

Acetone, AMG9810, dibutylphthalate (DBP) and dimethyl sulfoxide (DMSO) were obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Blasticidin, zeocin and tetracyclin were purchased from Life Technologies (Carlsbad, CA, USA). Dulbecco’s modified Eagle’s medium (DMEM) and HAM’s F12 were purchased from Nissui Pharmaceuticals (Tokyo, Japan). Ethanol was purchased from RCI Labscan Limited (Bangkok, Thailand). Fetal bovine serum (FBS) was obtained from Hyclone (South Logan, UT). Fluorescein-4-isothiocyanate (FITC), Fluo 4-AM and HEPES were obtained from Dojindo Laboratories (Kumamoto, Japan). Ionomycin was obtained from LKT Laboratories Inc. (St. Paul, MN, USA). L-Glutamine was purchased from Nacalai Tesque., INC. (Kyoto, Japan). Pentobarbital sodium was obtained from Kyoritsu Seiyaku Corporation (Tokyo, Japan). Piperine was purchased from TCI Co., LTD. (Tokyo, Japan). Probenecid and Poly-L-lysine were obtained from Sigma (St. Louis, MO, USA). EDTA was purchased from KANTO Chemical Co., INC. (Tokyo, Japan).

Capsaicin (Wako), menthol (Wako), bradykinin (Wako), ProTx-II (Wako), histamine (Wako), pyrilamine (Sigma), chloroquine (Wako), Nav1.7 inhibitor (GLPBIO), AMG9810 (Wako), PEAQX (GLPBIO) and NBQX (MedChemExpress) were dissolved in DMSO, ethanol or sterile, distilled water at the maximum concentration guaranteed by the manufacturer. The working concentration was prepared as 100 × dissolved in sterile, distilled water.

Rabbit anti-CLDN1, mouse anti-CLDN4, rabbit anti-ZO-1, Alexa Fluor 488 anti-mouse, and Alexa Fluor 555 anti-rabbit antibodies, and H₂DCFDA were obtained from Thermo Fisher Scientific (San Diego, CA, USA). Goat anti-β-actin antibody was from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse anti-phosphoserine (p-Ser) and anti-phosphothreonine (p-Thr) antibodies were from Sigma-Aldrich (Saint Louis, MO, USA). LY, NiSPY-3, Quest Fluo-8 AM, rabbit anti-nitrotyrosine, rabbit anti-NOS1, rabbit anti-NOS2, rabbit anti-NOS3, and rabbit anti-ubiquitine were from Biotium (Fremont, CA, USA), Goryo Kagaku (Hokkaido, Japan), AAT Bioquest (Sunnyvale, CA, USA), R&D Systems (Minneapolis, MN, USA), Cell Signaling Technology (Beverly, MA, USA), ProteinTech (Rosemont, IL, USA), GeneTex (Irvine, CA, USA), and Stressgen Biotechnologies Corporation (British Columbia, Canada), respectively. NOC12, SIN-1, and L-NAME were from Dojindo Laboratories (Kumamoto, Japan). AMG9810, olvanil, and RN1734 were from FUJIFILM Wako Pure Chemical Corporation (Tokyo, Japan). DAF-2DA and FeTPPs were from Cayman Chemical (Ann Arbor, MI, USA). All other reagents were of the highest grade of purity available.