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Effect of Cell Division on Cell Differentiation (RNA-Seq H9 cells UD E24 E24RO E24woTET E24TET)

ABSTRACT: Effect of Cell Division on Cell Differentiation (RNA-Seq H9 cells UD E24 E24RO E24woTET E24TET)

PROVIDER: PRJNA541528 | ENA |

REPOSITORIES: ENA

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			Action	DRS
		Other
	SRR9024860_1.fastq.gz	Fastqsanger.gz
	SRR9024860_2.fastq.gz	Fastqsanger.gz
	SRR9024862_1.fastq.gz	Fastqsanger.gz
	SRR9024862_2.fastq.gz	Fastqsanger.gz

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Project description: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.

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Project description:We addressed the role of cell cycle regulator CDK1 during differentiation by either inhibiting its activity with the specific inhibitor RO3306 (10µM) or by knocking its expression down using specific shRNAs, during mesendoderm differentiation. Human pluripotent stem cells (H9 cells) were synchronized in their cell cycle and then induced to differentiate into mesendoderm for 24h. shRNAs against CDK1 were induced 4 days before starting the differentiation. The CDK1 inhibitor was added to the medium 8h after the induction of differentiation. At the end of the experiment, the cells were harvested and processed for RNA extraction.

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