Effect of mutations in Lhx3 on interactions with Ldb1 and Isl1.

<p>Yeast two-hybrid data for Lhx3 (constructs comprised both LIM domains), the Lhx3 phosphomimic mutants Lhx3(S71D) and Lhx3(S71E) and the CPHDS mutation Lhx3(Y114C) against Ldb1_LID and Isl1_LBD, and. Interaction data for both bait/prey orientations are shown; DBD and AD designate the plasmids pGBT9 and pGAD10, respectively. Serial dilutions of culture (A_600nm = 0.2, 0.02, and 0.002) were spotted onto each column of the plate with the highest concentration at the top. Two different selection conditions (moderate stringency = SD -L-W-H +3-AT; high stringency = SD-L-W-H-A). Data are representative of 2­ or 3 separate experiments. In all cases transformation control plates (SD-L-W) showed strong growth of yeast indicating successful transformations, and negative controls of each constructs versus the corresponding empty plasmid showed no yeast growth under the conditions tested.