Protocol for chromatin immunoprecipitation and hematovascular function assays with differentiated mouse embryonic stem cells

Here, we present a protocol for mouse embryonic stem cells (mESCs) as a valuable model for studying developmental processes. This protocol details step-by-step differentiation of mESCs into embryoid bodies (EBs), followed by single-cell dissociation and chromatin preparation for chromatin immunoprecipitation (ChIP), including crosslinking and sonication. We also describe procedures to assess hematovascular differentiation including EB vascular sprouting assay, hematopoietic colony formation assay, and flow cytometry. This protocol enables the investigation of the dynamic interplay between epigenetic modifications and hematovascular lineage commitment during mESC differentiation. For complete details on the use and execution of this protocol, please refer to Kim et al.¹ and Park et al.².