Early anteroposterior regionalisation of human neural crest is shaped by a pro-mesodermal factor

Undifferentiated human embryonic stem cells (hESCs) were treated with WNT and FGF agonists for three days to generate NMP-like axial progenitors (NMPs). Overall design: Triplicate samples of undifferentiated hESCs (TBXT NEGATIVE) and NMPs (TBXT POSITIVE) were harvested, pooled and analysed by ChiP-seq We employed ATAC-seq to interrogate the chromatin accessibility landscape in neuromesodermal progenitor (NMP)-like cells and their derivatives, trunk neural crest and pre-neural spinal cord progenitors following differentiation of a human embryonic stem cell (hESC) line engineered to exhibit shRNA-mediated, tetracycline (Tet)-inducible knockdown of TBXT expression. hESCs were first differentiated towards NMPs (Day 3) and subsequently either trunk neural crest (tNC; upon culture in BMP/WNT agonists for 5 days) or pre-neural spinal cord progenitors (PNE; upon culture in WNT/high FGF for 4 days) in the presence (Pos) or absence (Neg) of Tetracycline.