ChIP-seq experiments to investigate the role of HOXB1 overexpression in cardiac progenitors
ChIP-seq experiments to investigate the role of HOXB1 overexpression in cardiac progenitors
Cells were fixed with 1% HCHO for 10 minutes, neutralised with 0.125M Glycine for 5 min, washed twice with ice-cold PBS and spinned down. Cell pellets were snap frozen and stored at -80C. Embryonic stem (ES) cells were cultured on a monolayer of mitomycin inactivated mouse embryonic fibroblasts and maintained in DMEM/F12-based medium containing 20% knockout serum replacement (Invitrogen) and 10 ng/ml bFGF (Miltenyi Biotech). hPSCs were passaged using TrypLE Select (Invitrogen). HOXB1_H11 cells were differentiated using the monolayer method as described in https://doi.org/10.1038/nmeth.1740. Doxycycline was added at day 4 of cardiac differentiation at 1 microgram/ml concentration. ChIPmentation samples and libraries were generated as described previously (https://www.doi.org/10.1016/j.xpro.2020.100187) with the use of the following antibody monoclonal ANTI-FLAG é€Ÿ M2 antibody (Sigma-Aldrich) Purified DNA was amplified using Nextera indexes following manufacturer's protocol.