Fig. 5: Transcriptional analysis of LC-PUFA-phospholipid biosynthetic pathways in the pregnant liver. | Nature Communications

Fig. 5: Transcriptional analysis of LC-PUFA-phospholipid biosynthetic pathways in the pregnant liver.

From: A co-ordinated transcriptional programme in the maternal liver supplies long chain polyunsaturated fatty acids to the conceptus using phospholipids

Fig. 5

A Schematic depicting the re-analysis of a published whole-transcriptome microarray dataset of livers from pregnant (14.5 dpc) and virgin mice (n = 4 mice per condition. B Heatmap showing z-transformed values of 3272 differentially expressed genes (DEGs) based on a p-value (corrected for multiple hypothesis testing based on the Benjamini–Hochberg procedure) threshold of 0.05 and a fold change threshold of 1.25. Differential expression analysis was performed using limma. List of DEGs is found in Supplementary Data Table S9. C Volcano plot showing DEGs (red) annotated with biosynthetic genes of interest. D Diagrammatic summary of hepatic lipid biosynthetic pathways chosen for targeted transcriptomics analysis annotated with microarray expression data (orange box = DEG that increased in pregnant livers; blue box = DEG that decreased in pregnant livers; grey box = gene not identified as differentially expressed). Known rate-limiting steps are indicated by thick black borders. Gene symbols are defined in Supplementary Data Table S10. E Schematic depicting real-time quantitative PCR (RT-qPCR) validation of candidate gene expression in livers from two genotype-matched virgin vs pregnant group comparisons. F–I RT-qPCR data is split into fatty acid import and synthetic pathway genes (F), Kennedy pathway genes (G), Land’s Cycle genes (H) and lipid export pathway genes (I). RT-qPCR data was normalised to housekeeping gene expression (Tuba1, Tbp and Hprt) and is shown as mean relative expression ± SD. Groups were called significantly different by two-tailed Mann-Whitney U tests (*p-value < 0.05; **p-value < 0.01; ***p-value < 0.001). p-values for Cd36: 1vs2 = 0.0003, 3vs5 = 0.0002; Fads1: 1vs2 = 0.0003, 3vs5 = 0.0002; Fads2: 1vs2 = 0.0006, 3vs5 = 0.0003; Elovl2: 1vs2 = 0.0041; Elovl5: 1vs2 = 0.0012, 3vs5 = 0.0006; Scd1: 1vs2 = 0.0012, 3vs5 = 0.0012; Gpam: 1vs2 = 0.0205 0.007; Agpat2: 1vs2 = 0.0003, 3vs5 = 0.0003; Agpat3: 1vs2 = 0.0003, 3vs5 = 0.0002; Pcyt1a: 1vs2 = 0.0006, 3vs5 = 0.0002; Pemt: 3vs5 = 0.0003; Lpcat3: 1vs2 = 0.0205; Pnpla3: 1vs2 = 0.0006, 3vs5 = 0.0002; Mttp: 1vs2 = 0.0035, 3vs5 = 0.0003; Apob: 1vs2 = 0.0003, 3vs5 = 0.0003; Lipc: 1vs2 = 0.0003, 3vs5 = 0.0003. n = 8 (group 1; exceptions: n = 7 for Scd1, Elovl2, Fads2, Chka, Pcyt1a, Mttp, Pnpla3), n = 7 (group 2; exceptions: n = 6 for Scd1), n = 8 (group 3; exceptions: n = 7 for Scd1, Fasn, Lipc, Elovl5, Agpat2, Lpcat2, Lpcat3), n = 8 (group 5; exceptions: n = 7 for Elovl5, Fads2, Etnk1, Pemt, Lpcat2; n = 6 for Scd1); mice per group. DG diglyceride, LPA, lyso-phosphatidic acid, LPC lyso-phosphatidylcholine, LPE lyso-phosphatidylethanolamine, PA phosphatidic acid, PC phosphatidylcholine, PE phosphatidylethanolamine, TG triglyceride. Source data are provided as a Source Data file.

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