GSM7993721: RELA_NT_rep1; Homo sapiens; OTHER
Formaldehyde (37%, Sigma-Aldrich, 25259) was directly added to the medium of the RPTEC to achieve a final concentration of 0.5% for 1 min in room temperature. Fixation reaction was quenched by adding glycine at a final concentration of 125 mM. The cells were treated with trypsin (Gibco, 0.05%) for 2 min at 37°C, scraped from a culture dish and centrifuged at 500 × g for 5 min. The centrifugated cells were resuspended in PBS with 1% BSA and counted. manufacturer's instructions for cross-linked samples. Library preparation was performed using the 500,000 cells in 100 ul wash buffer were mixed and incubated with Concanavalin A (ConA) conjugated paramagnetic beads. Antibodies were added to each sample (RELA antibody [Santa Cruz, sc-109, 1:25] or rabbit IgG negative control antibody [Epicypher, 13-0041, 1:50]). The remaining steps were performed according to the manufacturer's instructions for cross-linked samples. Library preparation was performed using the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England BioLabs, E7645S) with the manufacturer's instruction, including minor modifications indicated by CUTANA kit.