TCR-stimulation induced Egr expressions were impaired in the <i>Zfat</i>-deficient DP thymocytes.

<p>(A)Immunoblots for Egr1, Egr2 and Egr3 before or at the indicated time points after the stimulation with plate-bound anti-CD3ε and anti-CD28 antibodies in DP cells from the indicated genotypes. Actin was used as a loading control. Data are representative of three independent experiments. (B) Immunoblots for Egr1, Egr2 and Egr3 in DP cells from the indicated genotypes before or at the indicated time points after the stimulation with plate-bound anti-CD3ε and anti-CD28 antibodies under the condition with U0126 in DMSO or with DMSO alone. Actin was used as a loading control. Data are representative of three independent experiments. (C, D) Quantitative RT-PCR analysis of <i>Egr1</i>, <i>Egr2</i> and <i>Egr3</i> expression before (C) or at the indicated time points after the stimulation with anti-CD3ε and anti-CD28 antibodies (D) in DP cells from <i>Zfat</i>^f/f and <i>Zfat</i>^f/f-<i>LckCre</i> mice. The relative expression for each gene was normalized by the expression of <i>Actb</i>. The results are presented as the value relative to the unstimulated DP cells from <i>Zfat</i>^f/f mice. The data are the mean ± s.d.; n.s., not significant.