The RNA helicase DDX6 regulates self-renewal and differentiation of human and mouse stem cells [polysome RNA-seq]

Analysis of adult and embryonci stem cells depleted for DDX6. Results provide insight into the regulation of human stem cell self-renewal and differentiation by the RNA-binding protein DDX6. Polysome cells were treated with cycloheximide (CHX) at 100 µg/ml for 5 min at 37°C. And then resuspend in polysome lysis buffer, loaded on the sucrose gradient and centrifuged at 35,000 x g (Beckman, rotor SW41) at 4 °C for 3 h. 500 µl fraction were collected using a FC203B (Gilson) fraction collector. Fractions containing monosome(MONO), light polysome (LITE), medium polysome (MID), heavy polysome (HEAVY), and total polsome (POLY) were pooled.