RNA-seq of Muscle stem cells (WT and Nedd4L-cKO) and Myoblasts (WT)

Muscle stem cell quiescence is a complex and dynamic state. In this study we examined the role of the Ubiquitin Protease System in muscle stem cell function and quiescence. Using a genetic mouse model where Nedd4L is selectively deleted in myogenic cells, we characterized the effect of the loss of Nedd4L on muscle stem cells. With RNA-Seq we determined that the genetic deletion of Nedd4L resulted in the transition of the muscle stem cells out of deep quiescence and into Galert. Characterized by the upregulation of cell cycle genes and differentiation genes and loss of expression in numerous known quiescence genes. Nedd4L-cKO muscle stem cells are more prone to differentiation and fail to expand properly in vitro. To determine the effect of the genetic deletion of Nedd4L on the transcriptome of MuSCs, RNA-Seq was performed on 1000 freshly isolated MuSCs from Nedd4L-WT and Nedd4L-cKO mice. For the comparison of the transcriptome between MuSCs and myoblasts, 1000 freshly isolated MuSCs from WT mice were used for RNA-Seq and from the same mice cells were allowed to grow in culture for 3 days and then 1000 cells were collected to prepare RNA-Seq libraries