A transcriptomic study of myogenic differentiation under the overexpression of PPAR? by RNA-Seq
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To study the cellular and molecular function of PPAR? in skeletal muscle differentiation, we have generated inducible gain-of-function to overexpress PPAR? in C2C12 skeletal muscle cells. In order to identify PPAR? targets, RNA sequencing (RNA-seq) was used to evaluate and quantify of transcriptomes and expression patterns during myogenic differentiation under the overexpression of PPAR?. The formation of myotubes and the expression of muscle-specific myogenic genes such as MyoD and MyoG may be inhibited by any altered expression of PPAR?. Multiple genes and pathways were significantly involved in this process, including 11 genes such as Fndc9 and Slc14a1 with fundamental change of regulation modes, 9 genes of which were validated by the data of qRT-PCR. Our studies demonstrate that PPAR? would play critical roles on the skeletal muscle cells differentiation, mediating crosstalk among several pathways and transcription factors. Overall design: Inducible PPAR? C2C12 cells were differentiated for 5 days. Uninduced cells were used as controls. Experiments were performed in triplicate. Gene expression profiling analyses by high throughput sequencing were performed on mRNA samples isolated from the sense PPAR? C2C12 cells (PPAR?/+) and the wildtype cells on 0d and 5d.



