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. 2021 Jun 16;118(6):1466–1478. doi: 10.1093/cvr/cvab197

Figure 2.

Figure 2

Suppression of the Hippo pathway, predicted from the RNA-Seq data. (A) Circos map of the genes regulated by the YAP1 and TEAD TRs of the Hippo pathway. Connecting lines show common and unique genes and are coloured based on the status of fold change in gene expression in ACM vs Control samples. Blue indicates suppression and red activation of gene expression. (B) Pie charts showing the number of suppressed and activated genes for each TR obtained from IPA. (C) Heat map of the YAP-TEAD targets showing reduced transcript levels (blue) of the majority of the genes in this pathway, as opposed to increased transcript levels (red) in ACM as compared to control hearts. Each lane represents one sample. (D) Gene set enrichment analysis (GSEA) plot showing enrichment of the YAP1 targets among the suppressed genes. GSEA plots are shown with corresponding normalized enrichment score (NES) and the q value. (E–H) Immunoblotting and the corresponding quantitative data (normalized to GAPDH) showing reduced levels of YAP1, TAZ, TEAD1 isoforms, and ANLN proteins in the right (E and F) and left (G and H) ventricular ACM heart samples as compared to control hearts. P-values were calculated by unpaired t test. (I–P) Immunofluorescence staining of thin myocardial sections for YAP1 (I), TAZ (K), TEAD1 (M), and ANLN (O) of the Hippo pathway in the control and ACM hearts. Quantitative data showing the percentage of nuclei stained for the selected Hippo pathway proteins in the five control and five ACM hearts (J, L, N, and P). Between 13 000 and 40 000 nuclei per group were counted for this analysis and P values were determined by unpaired t test.