T-cell–enriched transcription factors. (A) A heatmap representation of enrichment scores for genes identified as highly enriched in T cells. Genes are ranked from highest to lowest T-cell score and top scoring BCL11B is the only TF highly enriched in T cells. (B) A histogram representation of the sum of enrichment scores in T cells for all genes. The top 2.5% genes of that distribution were accepted as significantly enriched and are shown in panel A. (C) Enrichment profile of ZBTB25 showing the top 20 enriched cells/tissues. (D) Effect of ZBTB25 depletion on TCR-stimulated NF-AT signaling. ZBTB25 knockdown Jurkat E-6 stable cells were electroporated with 8 μg of NF-AT–luc reporter and 1 ng of renilla-luc reporter. After 18 hours of electroporation, cells from each sample were dispensed into 3 equal aliquots with 1 mL of complete IMDM media with or without anti-CD3 plus anti-CD28 antibodies (1 μg/mL of each) or PMA (50 ng/mL) plus anti-CD28 antibody (1 μg/mL). After another 7 hours of incubation, cells were harvested and examined for luciferase activity. The experiment was done in triplicates. (E) Effect of ZBTB25 depletion on the induction of TCR stimulated NF-AT target genes. Indicated ZBTB25 knockdown Jurkat E-6 stable cells were incubated with or without anti-CD3 plus anti-CD28 antibodies (1 μg/mL of each) for 24 hours. Then 3 sets of 1 × 10⁶ cells from each experimental condition were harvested independently for real-time RT-PCR analysis of indicated gene expression. The experiment was done in biologic triplicates.