Egr2 is associated with DGK-α promoter upon anergy induction and can regulate DGK-α expression. (A) The homologous region of human and mouse DGK-α promoter sequences. The boxed site is a putative Egr2 binding site. The arrows represent the predicted transcriptional start sites of human and mouse DGK-α according to NCBI reference sequences. (B) pGL10 Th1 T cells were anergized with 1 µg/ml of immobilized anti-CD3, and Egr2 and DGK-α mRNA expression was assessed by qRT-PCR at the indicated time. (C) Jurkat cells were cotransfected with a control or a DGK-α reporter, and an EV or an Egr2-expressing vector, and luciferase activity was assessed 48 h later. (D) OVA-specific CAR Tg × Egr2^flox/flox Th1 T cells were untreated (Control) or anergized with immobilized anti-CD3 mAb (Anergic) for 6–8 h and immunoprecipitated with an anti-Egr2 or a control IgG Ab, followed by qPCR using specific primers for the DGK-α promoter region. Data are presented as mean ± SD, and are representative of two to three independent experiments. *, P < 0.05; **, P < 0.01.