FIG. 5.
DAX-1 domains involved in RNA binding. (A) Full-length DAX-1 and truncated proteins corresponding to aa 1 to 69 (NR1), 1 to 135 (NR1-R2), 1 to 204 (NR1-R3), and 205 to 470 (C, corresponding to the DAX-1 putative LBD) were translated in the rabbit reticulocyte system and tested in the RNA homopolymer binding assay. Lane I, 1:10 of the input protein. The assay was performed in a buffer containing 0.25 M NaCl. (B) RARγ LBD (aa 178 to 423) was translated in the rabbit reticulocyte system, and binding to RNA homopolymers was tested in the presence (lanes 6 to 9) and in the absence (lanes 2 to 5) of 1 μM all-trans-retinoic acid. Lane I, 1:10 of the input protein (lane 1). Bacterially expressed RARα was used in the RNA homopolymer binding assay and detected by Western blot, as described (12) (lanes 11 to 14). Lane I, 1:10 of the input protein (lane 10). The assay was performed in a buffer containing 0.25 M NaCl.