3C-seq for SAMD4A promoter for 30 min +DRB treatment upon 7SK insertion
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.  3C-seq was performed in duplicates as described (Stadhouders et al., 2013). In brief, ~10 million HUVEC nuclei were isolated at 0, 30, and 60 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. Ligated DNA was recut with DpnII and ligated, and PCR was performed for each viewpoint
Erasmus MC
3C-seq for SAMD4A promoter for 30 min +DRB treatment upon 7SK insertion