GSM9453019: Jurkat cells, IRF4_WT, Input; Homo sapiens; ChIP-Seq
For C91/PL cells, cross-linking was performed by adding 1/10 volume of freshly prepared formaldehyde solution (11% (v/v) formaldehyde, 100 mM NaCl, 1 mM EDTA (pH 8.0), and 50 mM HEPES (pH 7.9)) directly to the culture medium or PBS and incubating for 15 min at room temperature. Fixation was quenched by adding one-twentieth volume of 1.25 M glycine and incubating for 5 min at room temperature. Cells were then collected and washed twice with chilled PBS containing 0.5% (v/v) Igepal. The cell pellet was snap-frozen on dry ice. Chromatin extraction, fragmentation, antibody immunoprecipitation, and ChIP experiments were performed at Active Motif using antibodies against Tax (Tax-specific monoclonal antibody Lt-4), IRF4, RelA, RelB, and H3K27ac. For Jurkat cells overexpressing IRF4, ChIP was performed using the SimpleChIP Plus Enzymatic Chromatin IP Kit (Magnetic Beads) (Cell Signaling Technology, #9005) according to the manufacturer's instructions. Libraries were prepared from ChIP and input DNA by consecutive enzymatic steps of end-polishing, dA-tailing, adaptor ligation, and PCR amplification. Libraries were prepared from ChIP and input DNA using the DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) (Cell Signaling Technology, #56795) together with Multiplex Oligos for Illumina (Cell Signaling Technology, #47538) according to the manufacturer's instructions.