FIGURE 6.

In vivo assays of the physical interaction between Far11 and phosphatases Pph21, Pph22, and Pph3. A, SUS assay of the interaction between Far11 and phosphatases Pph21, Pph22, and Pph3 (see ”Experimental Procedures“ for details). The positive interactions are confirmed by the activation of the LACZ reporter. Far3 and Tpd3 were used as positive controls. Additional negative (−) and positive (+) internal controls were included as described (31). The SUS experiments were carried out with yeast strains transformed with either pESC-URA/CASP10 (upper panels) or the empty vector (lower panels) in galactose-containing medium. B, BiFC assay of Far11-VC-YFP and Pph3-VN-YFP. A Far3-VN-YFP was included as a positive control. The corresponding haploid strains harboring the VN-YFP fusions were used as negative controls (upper panels). The protein interactions were confirmed by the reconstitution of a functional YFP protein. The experiments were also carried out with the diploid strains transformed with either pESC-URA/CASP10 (middle panels) or the empty vector (lower panels) in galactose-containing medium.