T2C for 30 minutes (ApoI) replicate 1 in 7SKi cells
A pair of ZFNs designed to specifically target an intronic region downstream the SAMD4A TSS was custom-produced (Sigma-Aldrich). Plasmids carrying the ZFN open reading frames were in vitro transcribed to obtain the ZFN mRNAs according to the manufacturer’s instructions.  T2C was performed in duplicates as described ( Kolovos et al., 2014). In brief, ~10 million HUVEC nuclei were isolated at 0, 30 min post-stimulation, following cross-linking in 1% paraformaldehyde (10 min, 20°C; Electron Microscopy Science), chromatin was cut using ApoI or HindIII (New England Biolabs), and ligated in dilution under conditions that allowed most nuclei to remain intact.
Erasmus MC
T2C for 30 minutes (ApoI) replicate 1 in 7SKi cells