3C-seq for SAMD4A viewpoint for 30 minutes +DRB of TNFa treatment upon p300 inhibition
HUVECs from pooled donors were grown to 85-90% confluence in Endothelial Basal Medium 2-MV with supplements (EBM; Lonza)and harvested at 0, or 30 min post-stimulation; 50 μM C646 (a p300 inhibitor; Bowers et al., 2010) was added for 1 h before harvesting. 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 viewpoint for 30 minutes +DRB of TNFa treatment upon p300 inhibition