Extended Data Fig. 5: Regulation of TBX21 expression by CNS-12.
a, Analysis of chromatin accessibility of Tbx21 locus in murine CD4+ T cell by DNase I hypersensitive sites sequencing (DNase-seq). Cells were stimulated with 20 ng/ml phorbol myristate acetate (PMA) and 2 μM calcium ionophore (CaI) for 4 hours (GSE67451). b, Analysis of chromatin accessibility of Tbx21 locus in murine CD8+ T cells by ATAC-Seq. Cells were stimulated with 10 ng/ml PMA and 0.5 μM ionomycin (Iono) or left unstimulated (resting) for 2 hours with or without 2 μM cyclosporin A (CsA) (GSE93014). c, Analysis of chromatin accessibility, RNA polymerase II (pol II) and H3K27ac binding in TBX21 gene locus in human CD4+ and CD8+ T cells. ATAC-seq data of human naive CD4+ T cells with or without anti-CD3/CD28 for 1 day (Top, GSE161096). ATAC-seq data of human naïve CD8+ T cells with or without anti-CD3/CD28 for 2 days (Middle, GSE212699). ChIP-Seq analysis of Pol II and H3K27ac binding to the TBX21 gene locus in human T cells activated with anti-CD3/CD28 Dynabeads for 24 hours (Bottom, GSE183883). d, Analysis of NFATc2 binding to CNS-12 of Tbx21 in CD4+ T cells from WT and Stim1/2CD4 mice by ChIP-qPCR. T cells were stimulated with anti-CD3/CD28 and cultured for 5 days, then restimulated with 20 nM PMA and 1 μM ionomycin for 1 hour followed by ChIP-qPCR. e, ChIP-Seq analysis of p300, H3K27ac and JUNB binding to the Tbx21 gene locus in mouse CD4+ T cells. Murine CD4+ T cell stimulated with anti-CD3/CD28 antibodies for 5 days (Top, GSE207265) or with 20 ng/ml phorbol myristate acetate (PMA) and 2 μM calcium ionophore (CaI) for 4 hours (Bottom, GSE67443). The statistical significance of differences between ChIP-seq peaks of test and control groups was calculated using MACS2 v2.1.1., and for ATAC-seq peaks using the DESEQ2 package. ***P < 0.001; **P < 0.01; *P < 0.05.
