4C-seq of resting and anti-CD3/CD28-stimuated Jurkat cells

To investigate the interacting loci within the region containing rs5837875, we employed the circularized chromosome conformation capture (4C) assay to elucidate the spatial interactions and regulatory networks associated with rs5837875, utilizing this specific region as the bait in Jurkat cells. We performed 4C-seq on both resting and anti-CD3/CD28-stimulated Jurkat cells. Anti-CD28 and anti-CD3 antibodies were utilized to activate T cells as previously described (Simeonov et al. Nature, 2017). Briefly, the culture plate was coated with 10 µg/ml of anti-CD28 (TONBO Biosciences, 40-0289) for 12 hours at 4°C prior to cell inoculation. Jurkat or CD4+ T cells were then seeded with 10 µg/ml of anti-CD3 (40-0038). The cells were harvested 24 hours later for subsequent experimental analysis.