Fig 4.

Sos7 and Spc7 localization interdependencies. (A) (Left panel) Western blot analysis of Sos7-GFP immunoprecipitates from equal amounts of protein extracts from wild-type (wt) and spc7-23 cells grown at the indicated temperature. γ-Tubulin, loading control. (Middle panel) Live-cell images of Sos7-GFP in wt and spc7-23 cells grown at 25°C or incubated at 33°C for 6 h. (Right panel) Quantification of these fluorescence signals. The number of cells analyzed per strain and temperature was 30. (B) (Top) Serial dilution patch tests (10⁴ to 10¹ cells) of the indicated strains grown at 18°C for 8 days. (Bottom) Live-cell images of Spc7-GFP in wt and sos7-Δ7 cells grown at 18°C. Bar, 5 μm. (Right panel) Quantification of these fluorescence signals. The number of cells analyzed per strain was 30. (C) Serial dilution patch tests (10⁴ to 10¹ cells) of sos7^ts cells transformed with plasmids overexpressing spc7⁺. Transformants were grown under plasmid-selective conditions at permissive (25°C or 30°C) or restrictive (28 to 34°C) temperatures for 5 to 6 days.