Fig. 4: Identification of the nascent liver proteome by SORT-AC in ethanol-induced liver injury mouse model.

a Schematic of the acute alcohol gavage mouse model establishing. In the workflow, AAV carried TTR-Cre (100 μL of 1 × 10¹² vg/ml per mouse) was orbital injected into C57BL/6J mice on day 0 and administered high-concentration ethanol gavage (4 g/kg) every two days. Meanwhile, a low concentration of ethanol (5%) and AlkK (30 g/L) was dissolved into the drinking water. The liver samples were collected from the mice after 27 days. b The Red Oil O staining of the mouse liver. The mouse without ethanol administered was used as control. The red regions indicated the fat droplets in which ethanol group has significant fat droplets accumulation. Scale bar, 100 μm. c Quantification analysis of (b). Data are the mean ± s.d.; n  =  6 fluorescence images per condition. d The strategy of identifying EtOH-enriched proteins. e Hierarchical clustering of different indicated treatments. f Volcano plot of the proteins identified in both SORT-AC groups with or without acute alcohol gavage treatment. Statistical significance was assessed using a two-sided, empirical Bayes moderated t-test. The dashed lines were under the significance curve parameter of S₀ = 0.1 and a false discovery rate <0.05. The proteins significantly enriched were colored in blue and highlighted as EtOH Enrich. g Enrichment analysis of EtOH Enrich proteins that major participate pathways, and the protein interaction network involved in monocarboxylic acid metabolic process. The blue bars indicated the representative pathways related to alcohol metabolism. GO term enrichment was performed using a one-sided Fisher’s exact test. The resulting p-values were adjusted for multiple hypothesis testing using the Benjamini–Hochberg procedure. h The ratio of membrane and transmembrane proteins in EtOH Enrich. i The annotation of the subcellular localization of EtOH Enrich membrane proteins. j The heatmap analysis of EtOH Enriched proteins participated in transcription regulator, xenobiotic metabolism, stress responses, and lipid metabolism. Source data are provided as a Source Data file.