Defining key transcriptional regulators of extravillous trophoblast differentiation [ChIP-seq]

During early human pregnancy, extravillous trophoblasts (EVT) play a central role in placental anchorage and blood vessel remodeling. Despite the essential roles in pregnancy, key factors and mechanisms underlying EVT differentiation remain largely unknown. Here, we defined key transcription factors (TFs) by monitoring the dynamics of transcriptome and enhancer usage during EVT differentiation. We confirmed the requirements of the defined TFs and identified their action mechanisms. ChIP-seq of H3K27ac was performed in human trophoblast stem cells (TSC), EVT differentiating cells (day 3, EVT D3), and fully differentiated EVT (day 8, EVT D8). ATAC-seq and H3K4me3 ChIP-seq were performed in TSC and EVT D8. Time-course ChIP-seq of TFAP2C and DLX6 were performed in TSC, EVT D2, EVT D5, and EVT D8. To map genomic targets of DLX5 and ASCL2, ChIP-seq using native antibodies was conducted, and NRIP1 targets were identified by biotin-mediated ChIP-seq (bioChIP-seq). Inputs were sequenced for a control.