FIGURE 1.

Hhex yeast two-hybrid screening. A, a diagram is shown of Hhex fusion proteins containing the Gal4 DNA binding domain (DBD) in the N terminus. These proteins were tested as putative bait in a yeast two-hybrid assay. An asterisk indicates the protein used as the final bait in the screening. The Hhex protein showing functional domains is also shown above as a reference. N-ter, N-terminal domain; HD, homeodomain; C-ter, C-terminal domain. B, shown is immunoblotting using an anti-Gal4 antibody of extracts from the AH109 yeast transformed with the bait. The yeast-expressing plasmid, pGBKT7-T, expressing the SV40 large antigen T protein was used as a positive control. C, an overview is shown of the Hhex yeast two-hybrid screening by sequential transformation of bait and prey (VP16 library). VP16 library, E9.5-E10.5 mouse embryo library made in the pVP16; SD/−L/−W/−A/−H, synthetic dropout medium lacking leucine, tryptophan, adenine, and histidine; SD/−L/−W/−A/−H/+X-α-gal, synthetic dropout medium lacking leucine, tryptophan, adenine, and histidine, and containing 20 μg/ml of X-α-gal.