Fig. 3: Mlkl^Adi-KO mice alleviate mitochondrial dysfunction and reduce inflammatory signaling in visceral WAT under HFD conditions.

Experimental setup: Transcriptomic analysis was performed on visceral WAT collected 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. Differentially expressed genes (DEGs) were identified and subjected to pathway enrichment analyses. a Principal component analysis (PCA) plot of transcriptomic profiles for Mlkl^Adi-KO and WT mice under NCD and HFD conditions. b Hierarchical clustering heatmap of z-scores of FPKM values for DEGs across NCD- and HFD-fed Mlkl^Adi-KO and WT mice. c Volcano plot of DEGs comparing HFD-fed mice. Red and blue dots indicate significantly upregulated and downregulated DEGs, respectively. d Validation of RNAseq data with RT-qPCR (Mlkl, Wee1, Gst4, Cxcr4, Adipoq, Sgk1, Bckdhb). Results are expressed as mean ± SD. e Venn diagram showing the number of uniquely expressed genes in mice under HFD conditions. f Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of upregulated and downregulated DEGs in HFD-fed Mlkl^Adi-KO mice. g Heatmap of oxidative phosphorylation-related DEGs in HFD-fed mice. Statistical analysis: DEGs were identified using a false discovery rate (FDR) threshold of < 0.05. Volcano plot was generated using VolcanoseR, while pathway enrichment analysis was performed using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Heatmaps were created using Morpheus after applying log₂ transformation (log₂ [FPKM + 1]) and Z-score normalization. Statistical differences in the RT-qPCR were analyzed using unpaired t tests. Normality was verified with the Shapiro–Wilk test and homoscedasticity with the F-test. *p < 0.05; ***p < 0.001.