A specific E3 ligase/deubiquitinase pair modulates TBP protein levels during muscle differentiation

To study the function of E3 ligase Huwe1 knockdown on C2C12 proliferation and differentiation, we use mRNA-seq to compare gene expression profiles between control wild-type C2C12 myoblasts and two Huwe1 knockdown C2C12 cell lines generated by different shRNAs on differentiation Day 0 (proliferation medium) and Day 6 (differentiation medium for 6 days). To study the potential roles of Huwe1 in maintainance of muscle morphology, we use mRNA-seq to compare gene expression profiles between control and Huwe1 knockdown myotubes. To study whether downregulation of USP10 deubiquitinase is of functional significance during muscle differentiation, we compare gene expression profiles between control wildtype and USP10 overexpressing C2C12 cells on differentiation Day 6. Overall design: Control and Huwe1 knockdown C2C12 cells were collected on differentiation day 0, day 6 and day 8. Control and USP10 overexpressed C2C12 cells were collected on differentiation day 6. Subsequently, directional mRNA-seq expriments were performed with mRNAs purified from the collected cells.

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Li L, Martinez SS, Hu W, Liu Z, Tjian R