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. 2021 Jan 19;28(7):2045–2059. doi: 10.1038/s41418-020-00732-5

Fig. 6. Identification of the bona fide TEAD1 target genes in postmitotic CMs in mice.

Fig. 6

Volcano plots demonstrate differentially expressed genes (DEGs) between control and Tead1 igKO (a) or icKO (b) hearts. Genes changed with fold change (FC) ≥ 2 and FDR < 0.05 are considered significant. c Venn diagrams illustrate the overlap of downregulated (left panel, 646 genes) and upregulated DEGs (right panel, 726 genes) between Tead1 igKO and icKO hearts. d Venn diagram depicts the bona fide TEAD1 target genes (455 in total) in adult mouse CMs as revealed by the overlap between TEAD1 binding genes as identified by ChIP-seq and genes commonly downregulated (284 genes) and upregulated (171 genes) in both Tead1 igKO and icKO hearts. e Reactome pathway enrichment analysis of the TEAD1 bona fide target genes in the adult mouse heart. f Heat maps from RNA-seq data show 11 nDNA-encoded mitochondrial ETC genes that are significantly downregulated in both Tead1 igKO (left panel) and icKO hearts (right panel). g Integrative Genomics Viewer (IGV) tracks of Ndufab1 gene locus to show a representative RNA-seq of Tead1 adult heart KO (igKO and icKO, upper panels) and ChIP-seq of TEAD1, H3K4me3 or H3K27ac modifications (bottom panel) in adult mouse hearts. A pair of arrows points to the TEAD1 binding region in Ndufab1 gene locus with which the DNA fragment was cloned to a luciferase reporter vector. qRT-PCR analysis of expression of TEAD1 target mitochondrial ETC genes in Tead1 igKO (h) and icKO hearts (i) at mRNA level. Gene expression in control hearts was set to 1 (red dashed line). N = 6–12. *p < 0.05. j–m The downregulation of NDUFAB1 at protein level is confirmed in both Tead1 igKO (j) and icKO (l) hearts by Western blot. The band intensity of NDUFAB1 in Western blots of Tead1 igKO and icKO heart samples is quantified and plotted as shown in “k” and “m”, respectively. Protein expression in control hearts is set to 1 (red dashed line). N = 6. *p < 0.05. n Ndufab1 luciferase reporter genes containing WT or mutation of the TEAD1 DNA binding element as shown in panel “g” were co-transfected with TEAD1 expression plasmid for dual luciferase reporter assays. Co-transfection with pcDNA empty plasmid served as control and the reporter activity from this control group was set to 1 (red dashed line). N = 4–6. *p < 0.05.