3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt hoechst 33342

Gigantol was extracted from stems of Dendrobium draconis Rchb.f., as previously described [45 (link)] and dissolved in dimethylsulfoxide (DMSO) at the indicated working concentrations. 3-(4,5-Dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT), Hoechst 33342, propidium iodide (PI), bovine serum albumin (BSA), dimethyl sulfoxide (DMSO), cocktail protease inhibitor, hematoxylin, and eosin were purchased from Sigma chemical, Inc. (Chemical Express, Bangkok, Thailand). RPMI-1640 medium, DMEM, phosphate buffer saline (PBS), glutamine, penicillin, and streptomycin were purchased from Gibco company (Gibthai, Bangkok, Thailand). Primary antibodies against CD133, ALDH1A1, total AKT, phosphorylated AKT (Ser473), total STAT3, phosphorylated STAT3 (Ser727), and GAPDH, horseradish peroxidase labeled secondary antibodies, and RIPA lysis buffer were purchase from Cell Signaling Technology (Theera Trading, Bangkok, Thailand). Pentobarbital sodium injection was purchased from Ceva Sante Animal (VET AGRITECH, Nonthaburi, Thailand). 3,3′-Diaminobenzidine tetrahydrochloride hydrate was purchased from TCI Co., LTD (Chemical Express, Bangkok, Thailand). Primary antibodies of Ki-67 and α-SMA and matched secondary antibodies were purchased from DAKO (Medicare Supply, Bangkok, Thailand).

Decursinol angelate (Item no: 25212), (PubChem CID: 776123) was purchased from Cayman (Cayman Chemicals, Ann Arbor, Michigan, United States). DMEM, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Hoechst 33342, dichlorodihydrofluorescein diacetate (H₂DCFDA), acridine orange (AO), dimethylsulfoxide (DMSO) and mitotracker were purchased from Sigma (Sigma-Aldrich, St. Louis, MI, USA). Fetal bovine serum was purchased from Gibco, (Gibco-Thermo Fisher, Waltham, MA, USA). Details of the primary and secondary antibodies used are mentioned in Supplementary Table S1. All the other solvents used for the study were of highest grade and supplied by Sigma (Sigma-Aldrich).