HDAC Enzyme Activity in Alcohol-Treated MDDCs

Normal MDDCs were treated for 24 hrs. with 0.1 and 0.2% alcohol (n = 3) followed by NE-PER nuclear and cytoplasmic extraction (Thermo Scientific, Cat. #78835). Whole cell lysate extraction was performed for MDDCs from alcohol users (n = 6) and controls (n = 6). HDAC enzyme activity was measured with the HDAC assay kit following manufacture's protocol (Active Motif, Cat. #102209) and as previously described [14 (link),22 ]. HDAC enzyme activity was calculated across the groups and graph as pmoles/min/mg of protein. The fluorescence was measured in a Biotek plate reader (Winooski, VT), 360 nm excitation and 460 nm emission.