Lieber decarli liquid diet

All animal protocols were approved by the University of Florida Animal Care and Usage Committee and were conducted in compliance with their guidelines. Ctgf conditional knockouts (Ctgf^k/k) were previously published.^{14 (link),15 (link)} These mice carried two loxP sites flanking exon 4 of Ctgf (termed Ctgf^f) gene and one allele of the human ubiquitin C promoter (ubc)-Cre/ERT2 transgene. At 3-week-old age, Ctgf^f/f mice carrying ubc-Cre/ERT2 were given IP injection of the tamoxifen suspension (75 mg/kg body weight) over 5 days and the resulting Ctgf^k/k mice lost exon 4 of Ctgf in genotyping analysis using primers and PCR condition described previously.^{16 (link)} One month later, these mice were fed with the Lieber-DeCarli liquid diet (BioServ, Flemington, NJ) containing 1% ethanol for 2 days followed by 2% ethanol for 10 days. Carbon tetrachloride (CCl₄, 1 μL/g body weight) was injected through IP at 1 day before the end of experiment.

For surgical resection, wild-type mice (n = 35) were subjected to PH by excision of the median and left lateral liver lobes at their stem under aseptic conditions according to previous publication.^{17 (link)}For CCl₄ intoxication in combination with or without ethanol feeding, wild-type or mutant mice (8–10 week old) were subjected to moderate ethanol feeding using the Lieber-DeCarli liquid diet (BioServ, Flemington, NJ) containing 1% ethanol for 2 days followed by 2% ethanol for the duration of the experiment based on previous publication.^{18 (link)} Isocaloric maltose was administered to a pair-fed cohort. After that, 2% ethanol was fed for the remaining experiments. An average of 13.1 mL of the 2% ethanol-containing diet was consumed per day. Pair-fed mice were given an isocaloric diet in which ethanol calories were substituted with calories from maltose dextrin. Pair-fed animals received a diet volume equivalent to that of their ethanol-fed experimental counterparts on the previous day to ensure equivalent calories were consumed between groups. No differences were seen in final body weight between pair and ethanol-fed mice at any experimental time point. Ethanol-fed (n = 35) or pair-fed (n = 35) mice received a single acute dose of CCl₄ (1 μL/g body weight) prediluted 1:3 in olive oil and administered via intraperitoneal (IP) injection.