Necrostatin 1 nec 1 n9037

Tetrahydrobenzimidazole (TMQ) 0153 was synthetized from p-benzoquinone as previously described^{12 (link)}. 2′-deoxy-5-azacytidine (5-aza; A3656), 3-methyladenine (3-MA; M9281), bafilomycin (baf-A1; #B1793), N-acetyl-L-cysteine (NAC; LAA21), thapsigargin (TSG; T9033), PP242 (P0037), shikonin (SHK; S7576) and necrostatin-1 (Nec-1; N9037) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Chloroquine (CQ; NZ-51031-K200) was purchased from Enzo Life Science (ENZ-51031-0050). Tiron (SC-253669), Trolox (SC-200810), buthionine sulfoximine (BSO) (SC-200824) were obtained from Santa-Cruz Biotechnology (CA, USA). Hydrogen peroxide (H₂O₂) was purchased from Junsei Chemical (23150-0350) (Tokyo, Japan).

Rapamycin and MK-2206 were purchased from Selleckchem (Houston, TX, USA). 3-Methyladenine (3-MA) (M9281) and necrostatin-1 (Nec-1) (N9037) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pan-caspase inhibitor z-VAD-fmk was purchased from ApexBio (Houston, TX, USA). Primary antibodies anti-LC3 A/B, anti-ATG5, anti-ATG7, anti-Beclin-1, and anti-RIPK3 used for Western Blot were purchased from Cell Signaling Technology (Beverly, Mass, USA), anti-RIPK1 antibody used for Western Blot was purchased from Santa Cruz (Beverly, Mass, USA) and anti-GAPDH antibody was purchased from Kangchen biotech (Shanghai, China). Anti-RIPK1 and anti-RIPK3 antibodies used for immunohistochemistry staining were purchased from Proteintech Group (Rosemont, IL, USA).

Human lung adenocarcinoma cell lines A549 (CCL-185), H1299 (CRL-5803) and Vero African green monkey kidney cells (CCL-81) were obtained from American Type Culture Collection (ATCC, Manassas, VA). Cells were maintained in DMEM supplemented with 0.1mM non-essential amino acids, 5% fetal bovine serum, and penicillin-streptomycin (all from Invitrogen, Carlsbad, CA). Expression plasmid: pBABEpuro-EGFP-LC3 (Addgene, #22405) was provided by Jayanta Debnath (University of California, San Francisco). Chloroquine (CQ, C6628) and Necrostatin-1(Nec-1, N9037) were purchased from Sigma-Aldrich (St. Louis, MO). Staurosporin (STS, 11055682001) was purchased from Roche. Z-VAD-fmk (161401-82-7) was purchased from Merck. SiRNA directed against SQSTM1/p62 (NM_003900) was synthesized by GenePharma (Sense, GGAGCACGGAGGGAAAAGAtt; Antisense, UCUUUUCCCUCCGUGCUCCac). The siRNA targeting ATG7 (Invitrogen, HSS116182), BECN1 (Invitrogen, HSS112731), and negative control siRNA (Invitrogen, 12935400) were all purchased from Invitrogen Stealth RNAi collection.