Xenbase Image ID: 151126

Figure 5. Comparison of AR- and GR-HRE binding to the HRE1/1 + Fox and in presence of FoxA1. (A) Xenopus oocytes were injected with 0.46 ng FoxA1 mRNA mixed with increasing amount of mRNA coding for either AR, 0.3–1.2 ng, or GR, 2.3–7.8 ng, followed by 4 ng of HRE1/1 + Fox ssDNA injection (Figure 2A). The intranuclear AR and GR concentration plotted as a function of injected mRNA, the inset diagram shows an expanded version of the AR diagram. Error bars signify average deviation of double samples. (B) HR-HRE binding based on DMS methylation protection at the HRE1/1 + Fox and plotted as a function of nuclear concentration of indicated HRs. The arrows show the graphically determined apparent Kd, i.e. the receptor concentration at 50% saturation of the HRE site. The nuclear HRE concentration was 0.12 μM (analyzed as in Supplementary Figure S1A). (C) FoxA1-DNA binding from the same experiment as shown in Figure 5A and B. (D) AR-DNA binding as a function of total nuclear AR concentration is shown, 0.2–0.69 μM of nuclear AR was expressed together with ∼0.6 μM FoxA1 and then 10 ng of ssDNA containing HRE1/1 + Fox was injected resulting in 0.24 μM of nuclear HRE. (E) Experiment described in Figure 5D. AR-HRE1/1 + Fox binding depicted as Fb/Facc plotted as function of Cf (i.e. nuclear AR not specifically bound to DNA). The stippled arrow indicates the graphically determined Cf at 50% GR-HRE binding, i.e the apparent Kd. Image published in: Belikov S et al. (2016) © The Author(s) 2015. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license Larger Image Printer Friendly View

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