Fig. 3

Identification and evaluation of candidate activity enhancers. A Heatmap showing the clustering of Myh genes interactions in quadriceps and soleus. The color scale indicates the degree of correlation (blue, low correlation; red, high correlation); B Principal component analysis (PCA) of Myh genes interactions in quadriceps and soleus. Each point represents a sample. The percentages on each axis represent the percentages of variation explained by the principal components. The clustering and PCA were generated using Pearson correlation coefficients for every 1 Mb bin cis-interaction site after merging the 4C sites of the respective biological replicates; C A Venn diagram showing the number of common and unique genome-wide interactions site of the Myh genes in the quadriceps and soleus; D RNA-seq gene expression for the fast Myh genes; E Candidate enhancers of fast Myh genes in quadriceps and soleus. 4C-seq interaction profile (black), ChIP-seq profiles for H3K27ac (blue), and ATAC-seq (red). The vertical dashed red line indicates the 4C viewpoint. Light bars indicate candidate enhancer regions. The blue and red vertical lines below the ChIP-seq and ATAC-seq profiles indicate the peak; F The relative promoter activity of different Myh4 promoter regions was evaluated by luciferase reporter assay in H293 T cells. The pGL3-Basic was used as a control; G Dual-luciferase reporter assays to determine Myh4 candidate enhancer activity in H293 T cells. The pGL3-Myh4-pro3 was used as a control. Data are represented as mean ± SD of three independent experiments, and p-values are calculated using Student’s t-test (*P < 0.05; **P < 0.01; ***P < 0.001)