Identification of Akt2-responsive genes in the mouse model skeletal muscle cell line C2C12

Insulin selectively regulates cellular metabolic processes accompanied by activation of protein kinase Akt. We measured transcriptomic data in the mouse model skeletal muscle cell line C2C12 expressing photo-activatable Akt2 (PA-Akt2) probes upon light or insulin stimuli to reveal links between Akt activity and transcriptional control. We identified the genes that were quantitatively changed by each stimuli. Sampling procedure is as follows: After serum starvation for 14 hours, cells were stimulated with either light or insulin (100 nM) until predetermined sampling time points. At the collection time points, RNA from each sample was collected using an RNeasy kit (Qiagen). The quality of the RNA collected was evaluated according to the RNA integrity number (RIN) and found to be suitable for further sequencing. Standard Illumina protocols were used to generate 100-base pair paired-end read libraries that were sequenced on a HiSeq 2500 or HiSeq 3000 platform (Illumina).