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Figure 5


Fig. 5. Distribution of mid1-4GFP in other polarity mutants. (A) GFP fluorescence in live bud6{Delta} (AP1758), tea3{Delta} (AP1717), tea1{Delta} (AP1727) and pom1{Delta}tea3{Delta} (AP1737) strains expressing mid1-4GFP, grown at 25°C. Bar, 5 µm. (B) Distribution of mid1-4GFP spots on the cortex of wild-type, bud6{Delta}, tea3{Delta}, tea1{Delta}, pom1{Delta} and pom1{Delta}tea3{Delta} cells. Presence of one or more spots of mid1-4GFP at the medial (a), lateral (b) and tip (c) cortex was recorded in 300 to 400 cells from three to four independent experiments. Error bars give standard deviation (s.d.). (C) Localization of pom1-GFP in wild-type (AP1329) and tea1{Delta} cells (AP1760) grown at 25°C. In tea1{Delta} cells, pom1-GFP is still consistently detected at the cell tips and in the region of septum formation. Bar, 5 µm. (D) Septum position defects in polarity mutants. Cells were grown at 25°C and stained for septa with fluostain I. The percentage of abnormally placed or oriented septa was recorded in 400 to 800 septated cells from two to four independent experiments. Error bars give s.d. 1, wild type; 2, bud6{Delta}; 3, tea3{Delta}; 4, tea1{Delta}; 5, pom1{Delta}; 6, pom1{Delta}tea3{Delta}. (E) mid1-4GFP expression level in polarity mutants. Western blot analysis of Mid1p [FC418 strain (Celton-Morizur et al., 2004), first lane and left arrow] and mid1-4GFP expression levels (right arrow) in wild type (1), bud6{Delta} (2), tea3{Delta} (3), tea1{Delta} (4), pom1{Delta} (5) and pom1{Delta}tea3{Delta} (6) strains grown at 25°C using affinity purified anti-Mid1p antibody and TAT1 monoclonal antibody against tubulin.





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