FIG.2.

K88E mutant interacts more strongly with WT PITX2a. The indicated baits and the preys PITX2a K88E (A) or PITX2a T68P (B) or Pit-1 (C) were mated as described in the legend to Fig. 1 and plated on SD/−His/−Leu/−Trp plates with 0 to 10 mM 3-AT (−HIS) and/or SD/−Ade/−His/−Leu/−Trp plates (−HIS −ADE) to measure the strength of interaction. As in Fig. 1, β-galactosidase activity was also measured. The data represent results from four to six separate colonies obtained from two to three separate matings and represent the mean β-galactosidase activity (relative to that of the vector) ± the standard error of the mean. (D) Western blot of the bait constructs shown in panel C with anti-GAL4 DBD antibody. Full-length PITX2a is indicated. (E) Background activity of PITX2a, K88E, and T68P baits with the prey vector was determined as described in the legend to Fig. 1A. ND, not determined.