Effect of Cell Division on Cell Differentiation (ChIP-Seq HUvsNoc)

We addressed the role of cell division during differentiation by blocking the progression of the cell cycle during mesendoderm differentiation. Human pluripotent stem cells (H9 cells) were synchronized in their cell cycle and then induced to differentiate into mesendoderm for 24h. To block the cells in their S phase, we added Hydroxyurea (200 uM) at the beginning of the differentiation process. To block the cells in their G2/M phases, we added nocodazole (100 ng/ml) 8h after the induction of differentiation. At the end of the experiment, the cells were harvested and processed for RNA extraction. Four different conditions were analyzed: Undifferentiated (UD), Mesendoderm (E24), Mesendoderm + Hydroxyurea (E24HU), and Mesendoderm + nocodazole (E24Noc). Two biological replicates were generated for each condition.