FIGURE 3:

Further analysis of septin–septin interactions using the tripartite split-GFP system. Each of the essential septins (Cdc3, Cdc10, Cdc11, and Cdc12) was tagged with an N-terminal β10 tag and a five-residue linker and then combined by mating, as in Figure 1A, with each of the five mitotic septins tagged at their immediate C-terminal end with β11, as indicated schematically in the left-hand diagrams. Except in the cases in which both copies of the same septin were tagged (e.g., β10-Cdc10 and Cdc10-β11), a second (WT) copy of each tagged subunit is present but has been omitted for clarity. The resulting diploids were grown, induced with galactose, washed, and imaged as in Figure 1B. (A) Diploids 73–77, (B) diploids 66–70, (C) diploids 101–105, and (D) diploid strains 80–84. For β10-Shs1 examined in the same way, see Supplemental Figure S4. The fiducial marker for the septin collar was Cdc10-mCh expressed in the same cells, except in the cells expressing β10-Cdc10 and Cdc10-β11, for which the marker was Cdc11-mCh. (E) Quantification, as in Figure 1C, of the data shown in A–D.