Illumina HiSeq 4000 sequencing; Reduction of PRC2 activity supports survival of TNBC cancer cells to cytotoxic treatments.
G-2 cells were treated with CAF (312,5 ng/ml cyclophosphamide, 15,6 ng/ml doxorubicin and 312,5 ng/ml 5-FU) in 15 cm plates for 48 hours in biological triplicates. DNA-Protein complexes were cross-linked with 1% paraformaldehyde in PBS for 20 min at RT. After extraction, the chromatin was sonicated in a Bioruptor Pico (Diagenode) to generate 300-500 bp fragments. 20-30 Âµg DNA was used per Chromatin Immunoprecipitation reaction. Finally, the immunoprecipitated DNA was purified by phenol-chloroform extraction. ChIP-sequencing libraries were generated using the MicroPlex Library Preparation Kit v2 (Diagenode) along the manufacturers recommendations.  The size of the generated libraries was estimated on High Sensitivity DNA chip (Agilent) using a Bioanalyser 2100.