Fig. 5: Mlkl^Adi-KO mice exhibit reduced hepatic steatosis and distinct transcriptional profiles under HFD conditions.

Experimental setup: Transcriptomic analysis was performed on liver samples from Mlkl^Adi-KO and WT mice fed either a NCD (n = 4 per group) or an HFD (n = 5 per group) for 16 weeks. Triglycerides quantification was conducted on liver samples from WT and Mlkl^Adi-KO mice fed either NCD (n = 8 per group) or HFD (n = 8-9 per group) for 16 weeks. a Representative hematoxylin and eosin (H&E) staining of liver sections from Mlkl^Adi-KO and WT mice under NCD and HFD conditions. (Scale bar, 100 µm) (b) Intrahepatic triglycerides (TG) levels measured in Mlkl^Adi-KO and WT mice under NCD and HFD conditions. c PCA of RNA-seq data showing sample clustering based on diet and genotype. d Hierarchical clustering heatmap of differentially expressed genes (DEGs) in HFD- and NCD-fed mice. e Volcano plot of DEGs comparing HFD-fed mice. f Venn diagrams of DEGs unique to HFD-fed WT (2458 genes) and Mlkl^Adi-KO (1832 genes) livers. g KEGG pathway enrichment analysis of DEGs in Mlkl^Adi-KO livers under HFD conditions, showing enriched upregulated and downregulated pathways. h Validation of RNAseq data with RT-qPCR (Ip6k2, Lpin1). Statistical analysis: DEGs were identified using a false discovery rate (FDR) < 0.05. Pathway enrichment analysis was performed using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Results are shown as mean ± SEM. RT-qPCR data were analyzed using unpaired t tests. Normality was verified with the Shapiro-Wilk test and homoscedasticity with the F-test. *p < 0.05; **p < 0.01.