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. 2015 Feb;185(2):485–495. doi: 10.1016/j.ajpath.2014.09.021

Figure 3.

Figure 3

AP-1 signaling controls basal XT-1 promoter activity in nucleus pulposus (NP) cells. A: Effect of Fos overexpression on XT-1-wild-type (WT) promoter activity. Note the dose-dependent increase in XT-1 reporter activity with increasing concentrations of Fos. B: Jun overexpression induced XT-1 promoter activity in a dose-dependent manner. C: Cotransfection of Jun and Fos significantly increased the XT-1 promoter activity compared with their individual addition. D: DN-AP-1 diminished the activity of XT-1 promoter in a dose-dependent manner. E: DN-AP-1 has no effect on XT-1 reporter activity harboring a mutation in −730/−723 bp AP-1 motif. F: DN-AP-1 suppresses activity of the XT-1 promoter with deletion of −730/−723 bp AP-1 motif. G: Genomic chromatin immunoprecipitation analysis to detect AP-1 binding to XT-1 promoter in human NP cells. Pull down using Jun antibody clearly showed significant enrichment of binding to AP-1 motifs at −730/−723 bp and −684/−677 bp over IgG alone. Negative primers (Neg) showed little or no amplification for experimental and IgG control samples. Values shown are means ± SEM of three independent experiments. ∗P < 0.05.