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. 2012 Aug 3;153(10):5082–5089. doi: 10.1210/en.2012-1308

Fig. 4.

Fig. 4.

TR binds to the TRE in the IYD gene in vitro. A, Schematic diagram of the promoter region of the X. tropicalis IYD gene. The IYD TRE is shown as a white box with arrow, indicating the orientation of TRE. Black box shows the first exon. B, Comparison of the sequences of the TRE of the IYD gene and the mutated TRE (mTRE) to the consensus TRE. The TRE half-sites are shown in bold letters. The mutated nucleotides in the mTRE used for the gel mobility shift assay are underlined. C, The IYD TRE binds to TR/RXR heterodimers in vitro. A gel mobility shift assay was done with TR/RXR heterodimers translated in reticulocyte lysate in vitro and labeled X. laevis TRβ promoter TRE, a well-characterized TRE (51), in the presence or absence of 4-, 20-, or 100-fold of unlabeled wild-type or mutant IYD TRE as the competitor. Unprogrammed reticulocyte lysate (URL) was used as a negative control. The arrowhead indicates the complex of TRβ TRE with TR/RXR. Note that the complex was competed away efficiently by an excess of unlabeled wild-type IYD TRE but not mutated IYD TRE, indicating specific binding of the IYD TRE to TR/RXR.