Figure 5. UTX demethylase activity is required for H3K27me3 removal on muscle differentiation gene.

Primary myoblasts from female Utx^KI/KI or Utx^WT/WT mice were isolated, sorted by FACS, and expanded in vitro. Myogenic differentiation was induced for 24 hours. (A and B) RNA-Seq analysis was then performed to identify genes whose expression is modified in differentiating myoblasts from Utx^KI/KI mice compared with Utx^WT/WT mice. (A) A heatmap is shown for selected genes of different ontologies that are differentially expressed in the Utx^KI/KI myoblasts (P < 0.05). (B) GO analysis of the genes downregulated in myoblasts from the Utx^KI/KI mice shows highly significant enrichment of genes involved in muscle development and function. (C) ChIP analysis was performed to analyze the enrichment of UTX at the Myog, CKm, and Tnnc2 promoters. (D) Native-ChIP analysis was performed to determine H3K27me3 enrichment at the Myog, CKm, and Tnnc2 promoters. Values are presented as the average enrichment as a percentage of input ± SD. Statistical analysis was performed using an unpaired t test where *P < 0.05, n = 3.