Journal: Genes & Diseases

Article Title: 8-Chloroadenosine suppresses hepatocellular carcinoma progression via ADAR1/PPARγ axis-mediated lipid metabolism

doi: 10.1016/j.gendis.2025.101874

Figure Lengend Snippet: 8-Cl-Ado suppresses lipid metabolism in liver cancer by regulating gene expression. (A) The Venn diagram showing common differentially expressed genes (DEGs, p < 0.05) in HepG2 and Huh7 cells treated with 8-Cl-Ado for 24 h. (B) KEGG pathway enrichment analysis of common DEGs (top 10 pathways; 4 lipid metabolism-related pathways highlighted in red). (C) Effect of 8-Cl-Ado on lipid accumulation in HepG2 cells (Left: Representative images of oil red O staining; Right: Quantification of lipid droplet detection by oil red O staining). (D) Effect of 8-Cl-Ado on intracellular total cholesterol (CHO) and triglyceride (TG) levels in liver cancer cells. (E) Schematic diagram of CHO and fatty acid biosynthesis pathways. (F) The heatmap visualizing the expression patterns and significance (Log 2 (fold change)) of key genes involved in CHO biosynthesis and fatty acid metabolism ( n = 20) in 8-Cl-Ado-treated HepG2 and Huh7 cells. (G) Effect of 8-Cl-Ado on the expression levels of key genes involved in CHO biosynthesis and fatty acid metabolism in HepG2 cells ( n = 20). All experiments were performed with at least three biologically independent replicates ( n ≥ 3). Data were presented as mean ± standard error of the mean. Statistical significance was determined using Student's two-tailed t -test for two-group comparisons or one-way ANOVA for multiple groups. ∗∗∗ p < 0.001.

Article Snippet: The levels of cellular triglycerides and total cholesterol were investigated according to the manufacturer's instructions (Cat. No. 000180 and 000220; Nanjing Jiancheng Bioengineering Institute).

Techniques: Gene Expression, Staining, Expressing, Two Tailed Test

Journal: JCI Insight

Article Title: CRISPR screening identifies DTX4 governing alveolar macrophage cholesterol efflux in pulmonary alveolar proteinosis

doi: 10.1172/jci.insight.201129

Figure Lengend Snippet: ( A ) Western blot analysis of indicated proteins in differently treated THP-1–derived macrophages. GAPDH was used as a loading control. ( B ) Quantification of cholesterol efflux using the BODIPY-cholesterol assay ( n = 3 biological replicates). ( C and D ) Representative images ( C ) and quantification ( D ) of Oil Red O staining showing intracellular lipid droplet accumulation under different treatment conditions ( n = 3 biological replicates). Scale bars: 20 μm. ( E – G ) Flow cytometry analysis showing BODIPY fluorescence in THP-1–derived macrophages across treatment groups, including density plots ( E ), ridgeline plot ( F ), and quantification of BODIPY hi cell proportions ( G ) ( n = 3 biological replicates). ( H ) Confocal images of THP-1–derived macrophages stained with BODIPY 493/503 (green) to visualize neutral lipids ( n = 3 biological replicates); nuclei were stained with Hoechst 33342 (blue). Scale bars: 5 μm. ( I – K ) Quantification of intracellular free cholesterol ( I ), cholesteryl esters ( J ), and total cholesterol ( K ) in THP-1–derived macrophages under different treatments ( n = 3 biological replicates). Statistical analysis was performed using 1-way ANOVA followed by Tukey’s post hoc test for multiple comparisons (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: In brief, cells were labeled with BODIPY-cholesterol (catalog HY-125746, MedChemExpress) by incubation in labeling medium (DMEM) containing 1× BODIPY-cholesterol and 2 μg/mL Sandoz (catalog S9318, Sigma-Aldrich), an acetylcholinesterase inhibitor, for 1 hour.

Techniques: Western Blot, Derivative Assay, Control, Cholesterol Assay, Staining, Flow Cytometry, Fluorescence