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. 2001 Feb 1;29(3):629–637. doi: 10.1093/nar/29.3.629

Figure 5.

Figure 5

Yeast two-hybrid assay for the in vivo interaction of chHAT-1 or single missense mutants of it as compared to the leucine zipper motif with chCAF-1p48. The GAL4-based system and yeast S.cerevisiae were used for the two-hybrid study. (A) The in vivo interaction of chHAT-1 with chCAF-1p48 was performed on an SD drop-out plate lacking Leu and Trp (a) and an SD drop-out plate lacking Leu, Trp and His, but with the addition of 10 mM 3-AT (b). Qualitative lacZ assays for the in vivo interaction of the two proteins were performed on filters placed on YPD medium (c). (B) Quantitative assays for the in vivo interaction of the two proteins were performed with ONPG as a substrate. (C) Qualitative lacZ assays for the in vivo interaction of single missense mutants of chHAT-1 with chCAF-1p48 were performed on filters placed on YPD medium. The expression of proteins in combination with available vectors was verified by western blotting using anti-GAL4 antiserum as the primary antibodies (data not shown).