Repli-seq analysis with our without Tsix in EpiSCs

During differentiation and development, the precise and fine dynamics from euchromatin to facultative heterochromatin often remain unclear. We focused on Xist and its negative regulator Tsix as a paradigm to explore heterochromatin dynamics during local gene silencing. Using female epiblast stem cells (EpiSCs), we introduced facultative heterochromatin at the Xist promoter by inducing Tsix. Accompanied with Tsix induction, silencing of Xist expression was observed. In this study, we explored whether the silenced Xist via Tsix induction affects replication timing on the inactive X chromosome, examined by Repli-seq analysis.