Hdac1

Tetrahydrocurcumin, curcumin and calebin-A were provided by Sabinsa Corporation^® (Piscataway, NJ, USA). Sulfaphenazole, ketoconazole, α-naphthoflavone, quinidine, DMSO, PEG-400 and β-glucuronidase type IX A (β-glucuronidase) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Analytical-grade formic acid and HPLC-grade acetonitrile were purchased from Fisher Scientific. Ultrapure water from a Milli-Q^® system (Millipore, Billerica, MA, USA) was used for mobile phase. The antioxidant activity kit, cyclooxygenase-1 and -2 inhibitor screening kits, lipoxygenase inhibitor screening kit, HDAC1 and PCAF inhibitor screening kits were purchased from Cayman Chemical Company (Ann Arbor, MI, USA). Vivid^® CYP2C9 green screening kit, Vivid^® CYP3A4 green screening kit, Vivid^® CYP1A2 blue screening kit and Vivid^® CYP2D6 blue screening kit were bought from Life Technologies™ (Carlsbad, CA, USA). Human CXCL9/MIG, Human TNF-α, Human IFN-γ and Human CXCL10/IP-10 DuoSet^® ELISA kits, solutions and reagents were purchased from R & D Systems^® (Minneapolis, MN, USA). CellTiter 96^® AQueous One Solution Cell Proliferation Assay (MTS) was purchased from Promega (Madison, WI, USA). Heat-inactivated fetal bovine serum was purchased from Gibco^® by Life Technologies™ and Roswell Park Memorial Institute (RPMI) 1640 medium and Penicillin Streptomycin solution were purchased from Corning.

HDAC activity of class I HDACs (HDAC1, 2, 3 and 8) was measured with the fluorometric HDAC Activity Assay kit: HDAC1 (10011563, Cayman Chemical, Ann Arbor, MI), HDAC2, HDAC 3, and HDAC 8 (50062, 50073 and 50068, BPS Bioscience, San Diego, CA), according to the manufacturer’s instructions. HDAC activity of class II HDACs (HDAC4 and HDAC6) were measured with the fluorometric HDAC Activity Assay kit: HDAC4, HDAC 4 (50064 and 50076, BPS Bioscience, San Diego, CA), according to the manufacturer’s instructions.
Due to interference, concentrations of >30 mM were not used in the IC50 calculation. To account for any interference at lower concentrations, all baseline measurements of blank wells (no HDAC enzyme) were performed and subtracted with the various concentrations of diacetyl.