RNA-seq on Hu5/KD3 cells cultured with or without DAPT at proliferating stage. RNA-seq on Hu5/KD3 cells cultured with or without DAPT at proliferating stage

DAPT(a Notch inhibitor) promted myotube formation of human muscle progenitors. To clarify the Notch signal-responsive genes that inhibit or augment myotube formation, we examined the gene expression in Hu5/KD3 (immortalized human myoblast) treated with DAPT for three days by RNA-seq analysis. Hu5/KD3 cells were seeded 5×104 cells per collagenⅠcoated 24-well plate, cultured with DMEM (High Gluc.) containing 20% FBS, 2% UltroserG and supplemented with penicillin(100 units/mL), streptomycin (100 units/mL), Amphotericin B (250 ng/mL), L-Glu (2 mmol/L). Next day, started to treat with 0.1% DMSO or 10 μM DAPT in 10% FBS/DMEM. After three days treatment, total RNA was extracted as one sample from three wells by using RNeasy mini kit (QIAGEN). RNA libraries were prepared for sequencing using standard Illumina protocols