THP-1 monocyte-derived macrophages (50,000 cells) were treated with Deoxyribonuclease I (Worthington, LS002004) at 200 U/ml for 30 minutes to digest free-floating and dead cell DNA. After washing with Dulbecco's phosphate-buffered saline, cells were harvested using TrypLE (Gibco, 12604013). The cell pellet was then resuspended in ATAC-seq RSB (RSB; 10 mM Tris-HCl pH 7.5 (Invitrogen™, 15567-027), 10 mM NaCl (Invitrogen™, AM9759), and 3 mM MgCl2 (Invitrogen™, AM9530G) in Nuclease-Free Water (QIAGEN, 981103)) containing agents like NP40 (Roche, 11332473001), Tween-20 (Roche, 11332465001), and Digitonin (Promega, G9441), and incubated on ice for lysis. Following lysis, cells were washed in ATAC-seq RSB with Tween-20 and then centrifuged to isolate nuclei. The isolated nuclei were subjected to a transposition process using a specific mix (25 μl TD Tagment DNA Buffer (Illumina, 15027866), 2.5 μl TDE1 Tagment DNA Enzyme (Illumina, 15027865), 16.5 μl DPBS, 0.5 μl 1% digitonin, 0.5 μl 10% Tween-20, and 5 μl Nuclease-Free Water) and incubated at 37°C with shaking for 30 minutes. The DNA was then cleaned using the MinElute Reaction Cleanup Kit (QIAGEN, 28206). This was followed by a PCR amplification step using the NEBNext® High-Fidelity 2X PCR Master Mix (NEB, M0541L) with adapter primers. After determining and executing any additional amplification cycles, the library was purified using AMPure XP beads (Beckman, A63880) and stored at -80°C for future ATAC-seq analysis.