T2C for 0 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 0 minutes (ApoI) replicate 1 in 7SKi cells