F1258sp

Wild-type and Nogo-B KO mice were fed with a Lieber-DeCarli liquid diet (F1258SP, Bio-Serv, Flemington, NJ) containing 5% ethanol for 6 weeks. For controls, WT and Nogo-B KO mice were given a control diet (F1259SP, Bio-Serv), which matched ethanol-induced calorie with maltose dextrin. These diets were prepared daily and provided in the late afternoon before a 12-hour dark cycle. Mice were pair-fed with all diets. Nine hours before sacrifice, mice received a single intraperitoneal LPS injection (500ng/g body weight) to promote hepatic inflammation. Liver tissues and plasma were collected.

We used the binge-on-chronic NIAAA (Gao) model with 8-week-old
mice(Chu et al., 2021 (link)). Briefly,
male or female mice were acclimated to the Lieber-DeCarli liquid control
diet (F1259SP; Bio-Serv, Flemington, NJ) or gradually introduced to and
increased on the ethanol diet (5% ethanol-w/v; F1258SP; Bio-Serv) for 5 days
followed by further feeding with the liquid control (pair feeding, PF) or
ethanol diet (alcohol feeding, AF, 5% (v/v)) for an additional 10 days (day
15) or 45 days (day 50). On the last day of feeding, mice were also given
ethanol 5 g/kg or maltose dextran 9 g/kg by gavage, and sacrificed 9 hours
later. The volume of control diet given to mice was matched to the volume of
ethanol diet consumed. Alcohol fed groups were allowed free access to the 5%
(vol/vol) ethanol diet. Control mice were fed the isocaloric control diet
throughout the entire feeding period. Body weights were measured every other
day, and food intake was checked every day.

Male C57Bl/6 control and Shp^-/- mice (Wang et al., 2002 (link)) of 8-10 weeks of age were housed individually in an environmentally controlled room and maintained on a 24h light/dark (LD) cycle (lights on at 6 am to 6 pm) with free access to water and food. Mice were fed a control liquid diet for 5 days to acclimatize to the liquid diet and tube feeding regime. After the acclimation period, mice were given either a control diet (Bio-Serv, product #F1259SD) or 5% Lieber-DeCarli ethanol liquid diet (Bio-Serv, product #F1258SP) for 10 days with diets changed daily at 5 pm as described previously (Tsuchiya et al., 2015 (link)). On the 11^th day at 9 am, mice were gavaged with a single dose of maltose dextrin (Control, 9 g maltose dextrin per kg of body weight) or ethanol (5 g ethanol per kg of body weight), respectively. Nine hours after the binge, mice (n=3 per group per genotype) were euthanized and serum and liver collected every 6 hr over a 24h LD cycle at Zeitgeber time (ZT) ZT12, ZT18, ZT0 and ZT6. Both control and ethanol diets were provided throughout the sample collection period after the binge. A dim red light at intensity of 1 μmol/m²s was used to collect tissues in dark conditions. All experimental procedures were approved and performed according to the standards of the Institutional Animal Care and Use Committee (IACUC) and The University of Connecticut.

To examine the effect of HDAC6 overexpression in liver ECs on ALD, we used Cdh5-CreERT2 mice and the NIAAA model for generation of ALD with a slight modification (3.75 g/kg body weight ethanol gavage instead of 5 g/kg in the NIAAA model)[16 (link)]. Cdh5-CreERT2 mice were administrated with an adeno-associated virus 8 (AAV8)-EF1a-DIO-mKate2 (Vector Biolabs, Malvern, PA)(control, n=25) or AAV8-EFS-DIO-mHDAC6 vector (Vector Biolabs)(n=26) retro-orbitally at a dose of 2 × 10¹² GC/100 μL per mouse. Three weeks later, mice(16 Kate2 vs. 16 HDAC6) started to be fed with a 5% ethanol-liquid diet(F1258SP, Bio-Serv, Flemington, NJ) for 10 days. On the morning of Day 11, they received an oral gavage of ethanol and were sacrificed 9 hours later to collect blood and liver tissues. For a control group, Cdh5-CreERT2 mice(9 Kate2 vs. 10 HDAC6) were fed with a control liquid diet(F1259SP, Bio-Serv) for 10 days and were sacrificed for sample collection. All mice were approximately 4 months old at the time of sacrifice. We received liver specimens of ethanol-fed Sprague Dawley rats (the NIAAA model) from Dr. Varman Samuel(Yale University).
All animal experiments were approved by the Institutional Animal Care and Use Committee of the Veterans Affairs Connecticut Healthcare System(#YI0001) and performed in adherence with the National Institutes of Health Guidelines for the Use of Laboratory Animals.

Mice were randomized and individually housed and provided the alcohol-free control Lieber-DeCarli rodent liquid diet (product # F1259SP; Bio-Serv, Flemington, NJ) for 1 wk. After this period of acclimation, mice were randomly assigned to receive either the alcohol-containing liquid diet (product # F1258SP; Bio-Serv; n = 11) or to the control (no alcohol; n = 14) liquid diet. Control mice were pair-fed the same volume of an isocaloric and isonitrogenous liquid diet. Food consumption was assessed daily and fresh food was provided daily. The alcohol-fed group was acclimated to the alcohol-containing diet using an escalating step-wise protocol which included 1 day of 10%, and 2 days each of 16%, 22%, and 30% kcal from alcohol. Feeding was continued for a total of 24 wks. Whole-body fat mass was assessed noninvasively on conscious animals using a ¹H-NMR analyzer (Bruker LF90 Proton-NMR Minispec: Bruker Optics, Woodlands, TX) prior to the introduction of the alcohol-containing diet and at 6 wk intervals.