A-B. Quantification of β-galactosidase activity in wild-type and ssa1D, ssa2D, ssa1D ssa2D and ssa1D ssa3D ssa4D strains expressing FUS1::ubiYlacZ. EY3136, EY3137, EY3138, EY3141 and EY3148 (non-reverting ura3- derivatives) harboring the FUS1::ubiYlacZ plasmid pDL1460 were grown at room temperature and exposed to the indicated amount of α factor for 2 hours. Extracts were prepared as described in Materials and Methods. Mean +/- S.E. is shown. p-value asterisk key: ns (not significant) >0.05, * = <0.05, ** <0.01, ***,0.001, **** <0.0001. C-F. Streakouts of strains on YPD plates grown at room temperature. Strains: WT EY3136 (C), ssa1D EY3137 (D), ssa2D EY3138 (E), ssa1D ssa2D EY3141 (F). G-H. Duplicate halo assays for WT (G) and ssa1D ssa2D (H) strains grown at RT. K. Patch mating of ssa1D, ssa2D, ssa3D and ssa4D single, double and triple deletion mutants. Patches of wild type and mutant strains were grown on YEP-2% dextrose agar plates and then mated against lawns of MATa lys9 wild-type cells for the indicated time and then transferred to YNB 2% dextrose petri plates to select for prototrophs. F. RT, 6 hour mating for WT, ssa2D ssa2D. I. 30°C, 6 hour mating for WT, ssa1D, ssa2D, ssa4D. J. RT, 3 hour mating for WT, ssa1D ssa2D, ssa3D ssa4D, ssa1D ssa3D ssa4D, ssa2D ssa3D ssa4D, ssa1D ssa2D ssa3D.