Figure 4. Comparison of GR-HRE binding without or with FoxA1. (A) The DNA construct HRE1/1ΔFox with a triple-point mutated FoxA1 site. (B) Oocytes injected with GR mRNA, 1.4–7 ng mRNA, followed by injection of ssHRE1/1ΔFox, 4 ng, were analyzed as in Figure 1A, the diagram shows the GR-DNA binding monitored by DMS methylation protection of corresponding guanines in BE1 and BE2 at lower strand as a function of nuclear GR concentration. A Hill plot is for the same data is shown to the right. (C) Oocytes injected with increasing GR mRNA, 1.5–9.2 ng and constant FoxA1 mRNA, 0.35 ng, followed by 4 ng ssDNA HRE1/1 + Fox injection and analyzed as in 4B. The oocytes contained 0.12 μM nuclear FoxA1 based on [14C]-lysine labeled oocytes analyzed as described in Supplementary Figure S3C. (D) A Hill plot based on data in Figure 4C after introducing Facc = 0.59 in order to determine the Kd for the accessible HRE sites.
Image published in: Belikov S et al. (2016)
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