The T-box transcription factor Eomesodermin (Eomes) regulates the lineage-dependent expression of interferon γ (IFN-γ). We previously showed that Eomes promotes IFN-γ production and interacts with multiple conserved noncoding sequences (CNS) across the Ifng locus in mouse lymphoma BW5147 cells. In the present study, we investigated the transcriptional regulation of IFN-γ by the nuclear factor κB (NF-κB) subunit RelA and nuclear factor of activated T cells c2 (NFATc2, also known as NFAT1) in Eomes-transfected BW5147 cells. Eomes promoted the interaction of RelA and NFATc2 with the Ifng promoter and five CNS, including CNS-22 and CNS+30 upon stimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin (IM). The dual NF-κB and STAT3 inhibitor TPCA-1 moderately reduced the PMA- and IM-induced IFN-γ transcription in Eomes-transfected BW5147 cells. TPCA-1 interfered with RelA binding to the Ifng promoter, CNS-22 and CNS+30. Moreover, TPCA-1 reduced the interaction of Eomes or NFATc2 with the Ifng promoter and CNS+30. The present results indicate that Eomes promotes the interaction of RelA and NFATc2 with the Ifng promoter and multiple CNS across the Ifng locus in BW5147 cells.

T-box 转录因子 Eomesodermin (Eomes) 调节干扰素 γ (IFN-γ) 的谱系依赖性表达。我们之前表明，Eomes 促进 IFN-γ 的产生，并与小鼠淋巴瘤 BW5147 细胞中Ifng基因座的多个保守非编码序列（CNS）相互作用。在本研究中，我们研究了 Eomes 转染的 BW5147 细胞中核因子 κB (NF-κB) 亚基 RelA 和活化 T 细胞 c2 (NFATc2，也称为 NFAT1) 的核因子对 IFN-γ 的转录调控。Eomes 促进了 RelA 和 NFATc2 与Ifng 的相互作用启动子和五种 CNS，包括 CNS-22 和 CNS+30，在用佛波醇 12-肉豆蔻酸酯 13-乙酸酯 (PMA) 和离子霉素 (IM) 刺激后。在 Eomes 转染的 BW5147 细胞中，NF-κB 和 STAT3 双重抑制剂 TPCA-1 适度降低了 PMA 和 IM 诱导的 IFN-γ 转录。TPCA-1 干扰 RelA 与Ifng启动子、CNS-22 和 CNS+30 的结合。此外，TPCA-1 减少了 Eomes 或 NFATc2 与Ifng启动子和 CNS+30的相互作用。目前的结果表明，Eomes 促进 RelA 和 NFATc2 与Ifng启动子和多个 CNS 跨BW5147细胞中Ifng基因座的相互作用。