A 769662

Na_v1.7-transfected human embryonic kidney 293 (HEK293) cells were described previously [12 (link),36 (link),37 (link)]. Stock solutions of A769662 (LC Laboratories, Woburn, MA), Resveratrol (Cayman Chemicals, Ann Arbor, MI) and PT1 (Tocris, Minneapolis, MN) were made in DMSO and diluted in external solution bringing the final concentration of DMSO to <1%. For in vivo experiments, A769662 and PT1 were diluted in 100 mM PBS. Metformin was purchased from LKT Laboratories (St. Paul, MN) and stock solutions were made in double deionized water.

The fifty-five protein kinase inhibitors (PKIs) were purchased from following sources: BML-275, FR 180204, IKK16, GW 843682X, NSC 109555, NU7441, PD407824, PF 573228, SB 218078, TCS PIM-1-1, TCS PIM-1-4a, and TPCA-1 from Tocris Biosciences (Bristol, UK); indirubin-3′-monoxime and Ro-31-8220 from Calbiochem (San Diego, CA, USA); A-769662, bosutinib, chelerythrine, CP690550, fasudil, gefitinib, imatinib, nilotinib, PKC412, roscovitine, SNS-314, and tozasertib from LC Laboratories (Woburn, MA, USA); AT7867, AT9283, AZD1152, AZD1480, BI 2536, BIX 02189, CHIR-99021, CI-1040, CYC116, danusertib, enzastaurin, GDC-0879, INCB018424, JNJ-7706621, KU-55933, LY2228820, MLN8237, PD-0325901, PF-4708671, PLX-4032, PLX-4720, SB216763, SNS-032, SP600125, VX-702, Y-27632, and ZM447439 from Selleck Chemicals (Houston, TX, USA); U0126 from Promega (Madison, WI, USA); TBCA from Millipore (Burlington, MA, USA).
All TNBC cells in this study were obtained from the American Type Culture Collection (Manassas, VA, USA). The cultured cells were monitor by trypan blue cell counting as described previously [58 (link)].

Mouse C2C12 myoblasts were obtained from the American Type Culture Collection (ATCC, Rockville, MD). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), Dulbecco’s phosphate buffered saline (D-PBS), horse serum, penicillin, and streptomycin were from GIBCO (Grand Island, NY). Zileuton and LTB4 were from Tocris (Bristol, UK), and AmplexRed from Invitrogen (Waltham, MA). A769662 was obtained from LC laboratories (Woburn, MA). Antibodies were obtained as follows: GRP78, ATF3, CHOP, Akt and pAkt (Ser473) (Thr307) from Santa Cruz Biotechnology (Santa Cruz, CA); AMPK, pAMPK, pACC (Ser79) and ACC from Cell Signaling Technology (Danvers, MA); pIRS-1 (Ser307) and IRS-1 from Bioworld (Minneapolis, MN); 5-LO from Novusbio (Littleton, CO) and β-actin from Sigma-Aldrich (St. Louis, MO). 5-LO siRNA (sc-29597), AMPK1/2 siRNA (sc-45313) and control siRNA (sc-37007) were from Santa Cruz Biotechnology (Santa Cruz, CA). Oligonucleotide primers were from Bioneer Co. Ltd. (Daejeon, Korea). LTB4 kits were from Enzo Life Sciences (Seoul, Korea) and MyBioSource, Inc. (San Diego, CA). CysLTs kit and U75032 were from Cayman chemicals (Ann Arbor, MI). IL-6 and TNF-α ELISA kits were from BD Biosciences (San Diego, CA). All other reagents were from Sigma Chemicals (St. Louis, MO) unless indicated otherwise.

All chemical reagents were from Sigma-Aldrich (St. Louis, MO) unless otherwise stated. Arhalofenate acid (MBX-102 acid), the active form of arhalofenate, was used for in-vitro studies. Arhalofenate (MBX-102) was used for in-vivo studies. MSU crystals were prepared as described previously [22 (link)], suspended at 25 mg/mL in sterile, endotoxin-free phosphate-buffered saline (PBS), and verified to be free of detectable lipopolysaccharide contamination by Limulus lysate assay (Lonza, Walkersville, MD). A-769662 was from LC laboratories (Woburn, MA). Antibodies to phospho-AMPKα (Thr172) and total AMPKα (recognizing both AMPKα isoforms), SIRT1, TFAM, TRX1, TRX2, and TXNIP were from Cell Signaling Technology (Danvers, MA). Antibodies to pro-caspase-1 and cleaved caspase-1 (p10) were from Biovision (Milpitas, CA) and Santa Cruz Biotechnology (Santa Cruz, CA), respectively.