Fig. 5

Analysis of DNA binding of NFATc1/A-Bio in CTLs by ChIP seq assays. a Scheme of the Nfatc1 BAC transgene expressing tagged NFATc1/A-Bio protein in mice. The two promoters P1 and P2 and the 11 exons of the gene are indicated. b Immune blots of whole cell protein extracts from CTLs expressing only the E. coli biotin-ligase BirA, or BirA and NFATc1/A-Bio. The upper blot was incubated with an Ab directed against streptavidin, the lower blot with the NFATc1-specific mAb 7A6.The star indicates a non-specific band. c Motif analysis of NFATc1 binding sites in the 19 759 NFATc1/A peaks identified with a p-value of −5 by MACS. The letter size indicates the frequency of nucleotides within those sites (presented as ‘Bits’). d Peaks for NFATc1/A-Bio and NFATc2¹⁹ (p-value of −12 in MACS) within a distance of 100 kb to the 357 genes that were 2.5-fold differentially expressed in Nfatc1 ^−/− (Nfatc1 ^f/f x CD4-cre mice), compared to WT CTLs. 161 genes show both NFATc1/A-Bio and NFATc2 binding. e Binding of NFATc1/A-Bio and NFATc2¹⁹ to the prototypical NFAT target genes Ifng, Il2 and Il3 in CTLs + . Right, real-time PCR assays of Ifng, Il2 and Il3 RNA levels in WT, Nfatc1 ^−/− and Nfatc2 ^−/− CTL + cells, normalized by Actb and relative to naïve CD8⁺ T cells