Journal: Oncogenesis
Article Title: Targeting mitochondrial phosphatidylethanolamine alters mitochondrial metabolism and proliferation in hepatocellular carcinoma
doi: 10.1038/s41389-025-00593-y
Figure Lengend Snippet: A Schematic created using BioRender illustrating PISD enzyme reaction with phosphatidylserine (PS) to form CO 2 and phosphatidylethanolamine (PE). B Schematic created using BioRender showing generation of shScrambled (shSCR) and shPISD lentivirus in 293T cells and subsequent infection and gene silencing in HEPA1-6 cells. C The shPISD lentivirus reduced PISD mRNA abundance ( n = 2 per group) and altered the abundance of various genes involved in phospholipid metabolism ( n = 5 per group). D PISD activity was reduced in shPISD treated cells ( n = 3 per group). E , F Thin layer chromatography images indicating shPISD treated cells had reduced mitochondrial PE content ( E ) and densitometry quantification of thin layer chromatography images presented in E ( n = 3 per group) ( F ). G A Seahorse mitochondrial stress test reveals shPISD treated cells have reduced FCCP stimulated respiration rates ( n = 10 per group). H shPISD treated cells have elevated proton leak, calculated by subtracting oxygen consumption rates (OCR) during basal conditions from OCR during oligomycin (oligo) treated conditions ( n = 10 per group). I , J Western blot image indicating shPISD treated cells have reduced complex I and IV protein abundance ( I ) and densitometry quantification of Western blot images shown in I ( n = 6 per group) ( J ). K 1- 14 C oleate oxidation is reduced in shPISD treated cells ( n = 6 per group). L 1- 14 C lignoceric acid oxidation is increased in shPISD treated cells ( n = 6 per group). M PCA plot of untargeted metabolomics data illustrating that shPISD treated cells have an altered metabolome compared to shSCR treated cells ( n = 3 per group). N Volcano plot of untargeted metabolomics data illustrating differences in metabolites between shSCR and shPISD treated cells ( n = 3 per group). O Heatmap of select lipid metabolism related metabolites that were differentially expressed between shSCR and shPISD treated cells ( n = 3 per group). The data are presented as means ± S.D.
Article Snippet: 1- 14 C Lignoceric Acid was purchased from American Radiolabeled Chemicals (ARC-0805-50, Saint Louis, MO, USA).
Techniques: Infection, Activity Assay, Thin Layer Chromatography, Western Blot, Quantitative Proteomics
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