SH-SY5Y cells (CRL-2266, Lot 63724189, ATCC, Manassas, VA) were cultured in a 1:1 mixture of EMEM (ATCC) and F12K medium (10025-CV, Thermo Fisher Scientific) with 10% (vol/vol) fetal bovine serum (ATCC) and 1% penicillin and streptomycin. Cells were plated at a density of 1.5 × 106 in 25 cm2 canted neck flasks (Corning, Corning, NY). We modeled chronic intermittent exposure to ethanol (CIE) by using a pattern of 4 h exposure to 40 mM ethanol on 4 successive days, followed by 3 days without, repeated for 3 weeks (see Supplementary Figure 1). One hour after seeding, the cells were treated for 4 h with a “binge level” concentration of ethanol (40 mM) or left untreated, after which both sets of cells were washed with 4 mL 1X phosphate-buffered saline (PBS), and medium without ethanol was added. The 4 h ethanol exposure, wash and fresh medium was repeated Monday through Thursday for one pair of samples and Tuesday through Friday for a second. After washing the cells on Thursday (or Friday for the second pair), all dishes were split and plated
