Fig. 3: ABHD10 downregulation and S-palmitoylated PRDX5 upregulation associated with murine ALD.

a Male C57BL/6 J mice were randomly assigned to three experimental groups (n = 6/group): (i) the vehicle group treated for 6 weeks with olive oil, intubation, 1-week rest, and 3 weeks with olive oil; (ii) the EtOH (3w) group treated for 6 weeks with olive oil, intubation, one-week rest, and 3 final weeks with intragastric EtOH; and (iii) the CCl₄ (9w)+EtOH group treated for 6 weeks with CCl₄ (0.2 ml/kg), intubation, 1-week rest, and 3 weeks with a lower CCl₄ dose (0.1 ml/kg) and intragastric EtOH. Mice were sacrificed for experimental analyses. b, c Assessment of serum transaminase (ALT and AST) levels. d–f Hepatic steatosis was quantified by analyzing d hepatic triglyceride levels as well as e Oil Red O (ORO), and f Sirius Red staining in five random 200× fields of view. Scale bar = 200 μm. g Myeloperoxidase (MPO)-positive cells quantified from five random 200× fields of view. Scale bar = 200  μm. h Assessment of composite liver injury scores. i Liver tissue RNA was used for qPCR analyses of ABHD10. HPRT1 was used as a housekeeping control. j ABHD10 and S-palm-PRDX5 protein levels respectively densitometrically measured via standard immunoblotting and streptavidin pulldown-based immunoblotting of liver tissue lysates. Calnexin was used as an S-palmitoylated control, and β-actin was used as a loading control. k Elk-3 protein levels densitometrically measured via immunoblotting of liver tissue lysates. β-actin was used as a loading control. l Liver tissue RNA were used for qPCR analyses of the Elk-3 targets EGR1, FOS, and HIF1A. HPRT1 was used as a housekeeping control. Data presented as means with SDs. *P < 0.05, **P < 0.01 [one-way ANOVA].