Bulk RNA seq of mouse epiblast stem cells with Mixl1 differential timing expression

During mouse gastrulation, the formation of germ layers proceeds concurrently with the transition pluripotency state and the allocation of the multipotent epiblast cells to the germ layer derivatives. Lineage analysis and fate-mapping studies have revealed that groups of cells in different regions of the epiblast and the primitive streak of the gastrulating embryo are allocated to separate progenitors of the ectoderm, mesoderm and endoderm lineages three germ layers with descendants of each progenitor contributing to specific types of germ layer derivatives. Spatiotemporal transcriptomic analysis of the epiblast cell population has inferred that besides the allocation of the epiblast cells to progenitors of germ layers, a population of putative mesendoderm progenitors may be present in the gastrulating embryos. We converted ESCs to EpiSCs (Mixl1DOX) in vitro, in which induction by DOX enhanced the expression of Mixl1 . Mixl1DOX EpiSC expressed FLAG-Mixl1 transcript in response to Doxycycline (Dox) treatment, and generated FLAG-MIXL1 protein (30KDa) in embryoid bodies Embryoid bodies (EB) were collected on respective days aftertreating with /without Doxycycline. Doxycycline induction, where appropriate, was achieved by adding 1 μg/mL Doxycycline (Sigma) to the culture.