Cucurbitacin d

Cyto-ID autophagy detection kit was obtained from Enzo Life Sciences (Villeurbanne, France). Live and dead cell assay kit was purchased from Abcam (Cambridge, UK). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), bafilomycin A1 (BaF A1), chloroquine (CQ), 3-methyladenine (3-MA) formononectin, decursin and N-acetyl-L-cysteine (NAC) were from Sigma-Aldrich (St. Louis, MO, USA). Alexa fluor-488, Alexa fluor-594 and Lipofectamine reagent were obtained from Invitrogen (Garlsbad, CA, USA). Protein A/G plus-agarose beads and p62 siRNA were purchased cell signaling (Danvers, MA, USA). pBABE-puromCherry-EGFP-LC3B (addgene 22418) and EF.STAT3C.Ubc.GFP (addgene 24983) were from addgene (Cambridge, MA, USA). Nodakenin was purchased from ChemFaces (Wuhan, China) and cucurbitacin D was obtained from Extrasynthese (Genay, France).

Cucurbitacin D was purchased from Extrasynthese (Genay Cedex, France). DMSO and MTT were purchased from Sigma-Aldrich (St. Louis, MO, USA). Propidium iodide (PI) was purchased from Invitrogen (Carlsbad, CA, USA). Annexin V, Alexa Fluor 488 conjugate was obtained from Life Technologies (Eugene, OR, USA). The antibodies against cleaved caspase-8, -3, p-STAT3 (Try705), p-IκB (Ser32/36), p-NF-κB p65 (Ser536), pro-caspase-3, and total STAT3 were obtained from Cell Signaling (Danvers, MA, USA). The antibodies against IKK, PARP/p85, p-IKK, and total NF-κB were obtained from Santa Cruz Biotechnology (Dallas, Texas, USA). IκB antibody was obtained from Millipore. Tubulin antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA). ABC kit and diaminobenzidine tetrachloride (DAB) were obtained from Vector (Burlingame, CA, USA).