Journal: Genes & Diseases

Article Title: Lactate transporter MCT4 regulates the hub genes for lipid metabolism and inflammation to attenuate intracellular lipid accumulation in non-alcoholic fatty liver disease

doi: 10.1016/j.gendis.2025.101554

Figure Lengend Snippet: MCT4 regulates hub genes interactions to influence hepatic lipid metabolism. Subconfluent iHPx cells were infected with Ad-MCT4, AdR-siMCT4, or Ad-RFP, and stimulated with 0.05 mM oleic acid for 48 h. Total RNA was collected for RNA-sequencing analysis. (A) Venn Diagram of the differentially expressed genes (DEGs). (B) Gene set enrichment analysis (GSEA) of all expressed genes (MCT4/siMCT4). (C) Clustering analysis of FPKM values of 556 DEGs. (D) Venn diagram of partial DEGs verified by touchdown-quantitative PCR. (E) The network landscape generated by Cytoscape indicates a potential interaction relationship (colors indicate the importance of genes). (F) Scatter plot for Arg2 TPM of Normal ( n = 31) and NAFLD ( n = 112) in GSE162694 dataset (a). Unpaired, two-sided Mann–Whitney U test P -value are depicted in the plot, and the plot shows the medians (black line), standard deviation, and P -value. Correlation analysis of Arg2 level with Mct4 in human NAFL liver samples ( GSE167523 , n = 51) (b). r and P -value were obtained via two-tailed nonparametric Spearman's test, and the plot shows the linear regression line (black line) and r and P -value. (G) Immunohistochemical staining of ARG2 expression in liver samples in E. Representative positive stains are indicated with black arrows (100 × and 400 × ). (H) Subconfluent iHPx cells were infected with Ad-MCT4 or Ad-GFP, and stimulated with oleic acid, and total cell lysate was subjected to western blotting analysis to assess MCT4 and ARG2 expression at 72 h. Each assay condition was done in triplicate, and representative images were shown. (I) Immunohistochemical staining of MCT4 and ARG2 expression in liver tissues retrieved in . Representative positive stains are indicated with red arrows (400 × ). MCT4, monocarboxylate transporter 4; NAFLD, non-alcoholic fatty liver disease; NAFL, non-alcoholic fatty liver; FPKM, fragments per kilo base per million mapped reads; TPM, transcript per million; ARG2, arginase 2.

Article Snippet: The sections were deparaffinized and subjected to hematoxylin & eosin staining (Solarbio, Cat# G1120) and immunohistochemical staining as described., For immunohistochemical staining, the tissue sections were deparaffinized, rehydrated, antigen-retrieval treated, blocked, and incubated overnight with primary antibodies against ARG2 (1:50–1:500 dilution; Santa; Cat# sc-393496) and MCT4 (1:1000–1:4000 dilution; Proteintech; Cat# 22787-1-AP), followed by staining with biotin-labeled goat anti-rabbit IgG or anti-mouse IgG/streptavidin-HRP kit (SP Kit, PV-9000, ZSGB-Bio, China).

Techniques: Infection, RNA Sequencing, Real-time Polymerase Chain Reaction, Generated, MANN-WHITNEY, Standard Deviation, Two Tailed Test, Immunohistochemical staining, Staining, Expressing, Western Blot