Ulk1s757

Leflunomide and A77 1726 were kindly provided by Cinkate Corporation (Oak Park, IL). Brequinar sodium (BQR), a potent inhibitor of DHO-DHase, was kindly provided by Dupont Corporate (Wilmington, DE). PLX4720 was purchased from Selleck Chemicals Inc. (Houston, TX). SP600125, U0126, PD98059, and LY294002 were purchased from Cell Signaling Technology (Danvers, MA). Rapamycin was purchased from Cayman Laboratories (Ann Arbor, MI). Bafilomycin, colchicine metformin, 5Z-7-oxozeaenol, PF-4708671, and oligomycin were purchased from Sigma (St. Louis, MO). Anti-actin mAb and PPP were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Antibodies against p62, LC3, ULK1, AMPK, JNK, Jun, AKT, S6K1, S6 and their corresponding phospho-antibodies including ULK1^S555, ULK1^S757, AMPK^T172, AKT^S473, S6K1^T389, S6^S235/236, TAK1^T184/187 were purchased from Cell Signaling Technology (Danvers, MA). The expression vector encoding RFP-LC3 (pmRFP-LC3) was purchased from OriGene Technologies, Inc. (Rockville, MD).

Leflunomide and A77 1726 were kindly provided by Cinkate Corporation (Oak Park, IL). SP600125 was purchased from Cell Signaling Technology (Danvers, MA). Rapamycin was purchased from Cayman Laboratories (Ann Arbor, MI). Bafilomycin, colchicine, metformin, 5Z-7-oxozeaenol, PF-4708671, and oligomycin were purchased from Sigma (St. Louis, MO). Anti-actin mAb was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). Antibodies against LC3, ULK1, AMPK, AKT, S6K1, S6, ACC (acetyl-CoA carboxylase) and their corresponding phospho-antibodies including ULK1^S555, ULK1^S757, AMPK^T172, mTOR^S2448, AKT^S473, S6K1^T389, S6^S235/236, ACC^S79, and TAK1^T184/187 were purchased from Cell Signaling Technology (Danvers, MA). Anti-SOD1 antibody was kindly provided by Dr. Han-Xiang Deng (Northwestern University, Chicago). The SOD1-GFP and SOD1^G93A-GFP expression vectors were prepared by inserting a GFP gene downstream of SOD1 in a pcDNA3.1 vector. The expression vector encoding RFP-LC3 (pmRFP-LC3) was purchased from OriGene Technologies, Inc. (Rockville, MD). The NSC34 cell line was complete DMEM medium supplemented with 10% fetal bovine serum, streptomycin and penicillin, and L-glutamine.