Fig. 5: Identification of the core promoter of mice OX40 gene.
From: Critical role of OX40 in the expansion and survival of CD4 T-cell-derived double-negative T cells
a Nucleotide sequence of the OX40 promoter region. Potential regulatory elements identified according to the Transcription Factor Binding Sites database TRANSFAC are shown underlined and identified by the appropriate symbols. b Promoter-binding transcription factor (TF) profiling array assay of mice OX40 core promoter was performed. This is a competitive binding assay performed to identify promoter-bound TFs through comparisons of the results in the presence (with promoter) or absence (without promoter) of the mouse OX40 core promoter. If the OX40 promoter contains a TF-binding sequence, it will display a lower chemiluminescence activity. c Relative change of chemiluminescence activity in the absence (control) of the OX40 core promoter with presence (control + promoter) of the promoter. d Relative mRNA levels of the transcription factors in the presence or absence of IL-2. The data are presented as the mean ± SD, n = 3 in each group. *p < 0.05, **p < 0.01, NS no significance. e ChIP analysis of cDNT stimulated with IL-2 (50 ng/ml) for 48 h. Conventional PCR was performed to measure the relative levels of the antibody-bound DNA fragments. Soluble chromatin from the cDNT was immunoprecipitated with the PPARα, PPARγ antibody, or incubated with normal rabbit serum (IgG) for control purposes. The amplification for soluble chromatin before immunoprecipitation are shown as input. f Real-time PCR was used to determine the levels of PPARα and PPARγ bound directly to OX40 promoter sequence (−945/−736 bp and −1017/−950 bp upstream from transcription start site). The data are presented as the mean ± SD, n = 5 in each group. *p < 0.05, NS no significance
