212 Chapter 8 Figure 2: Identification of differentially expressed genes between PLN-R14del and control hearts using RNAseq. A) An overview illustrating the study design of comparing transcriptome between PLN-R14del and control hearts using RNA-seq after poly(A) selection. B) Heatmap showing top 100 differentially expressed genes in cardiac tissue from PLN-R14del versus control hearts. C) Examples of 3 differentially expressed genes between PLN-R14del and control hearts. D) PLN-R14del heart showed lower ATP2A2/SERCA2A signal (purple) and HADHA signal (red) compared to the control heart using immunofluorescence staining, confirming its levels in the RNAseq data. Nuclei were stained by DAPI (blue) and sarcomeres were stained by TNNI3 (green). Confocal images were taken at 63x magnification. E) Enrichment analysis using downregulated genes indicated the top enriched biological processes and pathways were related to metabolism. F) Protein-protein-interaction network using 39 TFs from the integrative analysis pointed toward (negatively) affected metabolism. Protein-coding genes involved in “Negative regulation of cellular metabolic process” are shown in red, protein-coding genes involved in “Cellular response to lipid” are shown in purple, and protein-coding genes that are not involved in these two processes are shown in light grey.
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