ChIPseq of RNA Exosome components in BMEL cell lines invalidated or not for the expression of the three HP1 isoforms (CBX1, CBX3, CBX5)
ChIPseq of RNA Exosome components in BMEL cell lines invalidated or not for the expression of the three HP1 isoforms (CBX1, CBX3, CBX5)
BMEL cells were crosslinked by a double procedure of 2 mM Di-Succinimidyl-Glutarate (DSG, Merck) and 1% formaldehyde. Chromatin was sonicated for 15 cycles of 30 seconds on and 30 seconds off. Following extraction, chromatin immunoprecipitation (ChIP) was incubated overnight with 4 µg of antibody. followed by the addition of protein G beads (Dynabeads) incubated for an additional 1h. Beads were then washed 5 times for 5 min each on a rotating wheel. Samples were incubated overnight at 65°C to reverse crosslink, followed by proteinase K and RNase A treatment. DNA was isolated using a NucleoSpin PCR purification kit (Macherey Nagel). ChIP-seq libraries were prepared with DNA Library Prep Kit for Illumina by the Genom'IC core facility (Paris, France).