Penicillin streptomycin antibiotic mixture

Jurkat and THP-1 cells were procured from ATCC. Epstein Barr Virus (EBV)-transformed B cell lines were obtained from the Lupus Family Registry and Repository (LFRR) housed by the Oklahoma Rheumatic Disease Research Cores Center (ORDRCC) at the Oklahoma Medical Research Foundation (OMRF) with IRB approval (22 (link)). Sanger sequencing was used to verify the genotype of EBV B cell lines carrying the risk (A) or non-risk (C) allele of the index SNP, rs140490. Jurkat, THP-1, and EBV B cells were maintained in RPMI 1640 medium supplemented with 10% FBS, 1X penicillin-streptomycin antibiotic mixture (Atlanta Biologicals, Inc.), and 2 mM L-glutamine (Lonza). THP-1 cell medium was also supplemented with 50 μM ß-mercaptoethanol. Where indicated, cells were stimulated with phorbol 12-myristate 13-acetate and ionomycin (P/I; 50 ng/mL, 500 ng/mL) for 2 h prior to harvest. For the siRNA knockdown of CTCF or YY1, EBV B cells homozygous for the UBE2L3-YDJC non-risk or risk haplotype were transiently transfected with 10 nM siRNA using 4D Amaxa Nucleofector Unit for EBV B cells (Lonza; Nucleofector SF kit, #V4XC-2032). The ON-TARGETplus human CTCF siRNA SMARTPool (#L-020165-00-0005), human YY1 siRNA SMARTPool (#L-011796-00-0005), and non-targeting scramble siRNA pool (#D-001810-10-05) were purchased from Dharmacon. All stock laboratory chemicals were from Sigma Aldrich or ThermoFischer.