GSM9883410: PC_CTL_ED_rep1; Drosophila melanogaster; OTHER
For experiments involving non-histone proteins, EDs were crosslinked with 0.1% formaldehyde for 5 min at RT on a rotating wheel. MNase digestion was performed using pAG-MNase Enzyme (Cell Signaling Technology) for 30 min at 4°C. Following reverse crosslinking for 4 h at 65°C and Proteinase K digestion, DNA was purified using SPRIselect beads and eluted in 50 μl of Tris-EDTA. The following antibodies were used: anti-H3K27me3 (1:100, Active Motif, catalogue no. 39155), anti-H2AK118ub (1:100, Cell Signaling Technology, catalogue no. 8240), anti-HA (1:50, Cell Signaling Technology, catalogue no. 3724) and anti-PC (SCHUETTENGRUBER et al. 2009). All experiments were performed in biological duplicates. DNA libraries for sequencing were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina. Paired-end sequencing (150 bp; approximatively 2 Gb per sample) was performed by Novogene (https://en.novogene.com/).