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. 2022 Oct 5;119(41):e2203480119. doi: 10.1073/pnas.2203480119

Fig. 2.

Fig. 2.

Restricting MUFAs activates the ER stress response pathway. (A) Volcano plot of changes in PC and SM (green), PE (blue), and TAG (red) relative peak intensities measured by lipidomics profiling analysis in shSCD vs. shCtrl OVCAR-5 cells cultured in low-serum conditions for 48 h. (B) Normalized peak intensities of PC lipids with differing fatty chain plasticity (0°, no carbon–carbon double bond; 1°, 1 or 2 carbon–carbon double bonds; 2°, more than 2 carbon–carbon double bonds) in shSCD vs. shCtrl OVCAR-5 cells. ShSCD cells have significantly fewer 1° PC lipids than shCtrl cells, whereas no difference was observed for 0° or 2° PC lipids. (C) Fold change of TAG lipids containing 16:0 or 18:0 fatty acyl chains integration and 16:1 or 18:1 fatty acyl chains integration determined by lipidomics profiling analysis in shSCD vs. shCtrl OVCAR-5 cells. (D) Volcano plot of changes in gene expression measured by RNA-seq in shSCD vs. shCtrl OVCAR-5 cells. Red dots represent up-regulated genes, and blue dots represent down-regulated genes. Genes of the ER stress response pathway are indicated by lines. (E) Dot plot of the top 15 enriched Gene Ontology terms identified by analysis of up-regulated genes (RNA-seq) in shSCD vs. shCtrl OVCAR-5 cells. Dot size corresponds to ratio of genes on the pathway vs. total number of up-regulated genes. Background color corresponds to FDR q value. (F) Enrichment plots generated by Gene Set Enrichment Analysis of gene expression (RNA-seq normalized counts) in OVCAR-5 shSCD vs. shCtrl using Hallmark and C2 gene sets from Molecular Signatures Database. (G) Top 10 significant upstream regulators identified by Ingenuity Pathway Analysis using DEGs from RNA-seq analysis of OVCAR-5 shSCD vs. shCtrl cells. Transcription factors involved in the ER stress response pathway are in bold characters. (H) Heat map of expression (RNA-seq normalized counts) and supervised hierarchical clustering of significant genes of the ER stress response pathway in shCtrl and shSCD OVCAR-5 cells (n = 3). *P < 0.05, ****P < 0.0001.