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. Author manuscript; available in PMC: 2014 Jan 1.

Published in final edited form as: J Immunother. 2013 Jan;36(1):3–10. doi: 10.1097/CJI.0b013e3182791234

For epigenetic analysis, two agents were used. (A) For methylation analysis, 5 representative clones were stimulated with OKT3/CD28 mAbs, in the presence of the methyltransferase inhibitor, 5-aza-2′-deoxycytidine (5-AzaC) at the indicated concentration. Four days after treatment, clones were analyzed for GFP expression by flow cytometry. (B) For histone deacetylation analysis, 5 representative clones were stimulated on OKT3/28 mAb-coated plates for 4 days. Six days after treatment, the histone deacetylase inhibitor, trichostatin A (TSA) was added into the cultures at the indicated concentration. Twenty-four hours after treatment, TSA-treated clones were analyzed for GFP expression by flow cytometry.

For epigenetic analysis, two agents were used. (A) For methylation analysis, 5 representative clones were stimulated with OKT3/CD28 mAbs, in the presence of the methyltransferase inhibitor, 5-aza-2′-deoxycytidine (5-AzaC) at the indicated concentration. Four days after treatment, clones were analyzed for GFP expression by flow cytometry. (B) For histone deacetylation analysis, 5 representative clones were stimulated on OKT3/28 mAb-coated plates for 4 days. Six days after treatment, the histone deacetylase inhibitor, trichostatin A (TSA) was added into the cultures at the indicated concentration. Twenty-four hours after treatment, TSA-treated clones were analyzed for GFP expression by flow cytometry.