Mocetinostat

Resveratrol, piceatannol, resminostat, entinostat, mocetinostat, vorinostat, curcumin, garcinol, anacardic acid and Tip60i were purchased from Selleckchem (Houston, TX, USA). MB-3 and BMS-345541 were purchased from MilliporeSigma (Burlington, MA, USA). Pterostilbene and myricetin were from LKT Laboratories (St. Paul, MN, USA) and trimethoxy-Resveratrol (trans-3,5,4′-trimethoxystilbene) was from Cayman Chemical Co. (Ann Arbor, MI, USA). Stock solutions of the chemicals were prepared based on the information provided by the manufacturer and maintained at −20°C. The antibodies for human PD-L1 (E1L3N, 13684), p38 MAPK (D13E1, 8690), NF-κB p65 (D14E12, 8242), γH2AX (20E3, 9718), cleaved caspase 3 (D3E9, 9579), IRF-1 (D5E4, 8478) and rabbit IgG isotype monoclonal antibody (DA1E, 5742) conjugated to PE were obtained from Cell Signaling Technology (Beverly, MA, USA). c-Myc antibody (9E10, sc-40) was obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Fetal bovine serum, RPMI-1640, DMEM, streptomycin and penicillin were from Gibco/Thermo Fisher Scientific (Waltham, MA, USA). All other chemicals and solvents were of analytical grade.

L3.6, BxPC3 and Panc1 cells were purchased from ATCC (Manassas, VA, USA) and cultured in their respective growth media supplemented with 10% fetal bovine serum (Sigma), 1× Pen/Strep (Sigma) at 37°C under 5% CO₂. L3.6 cells were grown in phenol red-free Minimum Essential Medium Eagle (MEM; Invitrogen) containing 1× l-glutamine, BxPC3 cells in phenol red-containing Roswell Park Memorial Institute (RPMI; Invitrogen) 1640 medium and Panc1 cells in phenol red-free Dulbecco's modified Eagle's medium/F-12 (DMEM/F-12; Invitrogen) growth medium. The authenticity of all cell lines used in this study was authenticated by DNA profiling using eight different and highly polymorphic short tandem repeat (STR) loci. Cells were treated with 4SC-202 (1 μM), mocetinostat (500 nM, Selleckchem), entinostat (500 nM, Selleckchem), JQ1 (250 nM) or DMSO (vehicle) for 24 or 72 h and TGFβ (5 ng/ml; R&D systems) for 72 h as indicated. siRNA transfections were performed using Lipofectamine^® RNAiMAX (Invitrogen) according to the manufacturer's instructions. SmartPool^® siRNA against MYC (Dharmacon) contained the following sequences: 5΄-AACGUUAGCUUCACCAACA-3΄, 5΄-GGAACUAUGACCUCGACUA-3΄, 5΄-GAACACACAACGUCUUGGA-3΄, 5΄-CUACCAGGCUGCGCGCAAA-3΄. siGENOME non-targeting siRNA (Dharmacon; D-001206-13) was used as a negative control.