Figure 4.

Single-residue substitutions/deletion within PalC conserved regions I and II largely impair or prevent its interaction with Vps32.A) Western blot analysis of yeast strains expressing the indicated fusion proteins, which were used as preys in two-hybrid assays. Fusion proteins were detected with anti-HA antibody. Actin was used as a loading control. B) PalC mutations affecting two-hybrid interaction with Vps32, as determined by quadruple selection on -Leu, -Trp, -His, -Ade medium using strain AH109. The experiment was carried out in triplicate. QSM indicates quadruple selection medium. -W-L is medium lacking Tp and Leu. C) PalC mutations affecting two-hybrid interaction with Vps32, as determined by quantitative β-galactosidase assays using strain Y187. Bars indicate standard errors.