Figure S2.
Role of the HSP40 Ydj1p, the HSP70 Ssa1p, and Rsp5p in regulating fMAPK activity. (A) Serial dilutions of wild-type cells (WT, PC538), ydj1Δ (PC7616), and ssa1Δ (PC7621) mutants grown on YEPD plates for 2 d. (B) Fluorescence microscopy of wild-type cells (WT, PC986) and the ydj1Δ mutant (PC7657) in the S288c background expressing GFP-Cdc42p (PC6454). Scale bar, 5 µm. (C) Fluorescence microscopy of wild-type cells (WT, PC6016) and the ssa1Δ mutant (PC7700) expressing GFP-Cdc42p (PC6454). Scale bar, 5 µm. (D) Microscopic examination of wild-type cells (WT, PC538), ydj1Δ (PC7616), tec1Δ (PC6102), and the tec1Δ ydj1Δ double mutants (PC7820) grown on YEPD plates for 16 h. Scale bar, 10 µm. (E) Microscopic examination of wild-type cells (WT, PC538), ssa1Δ (PC7621), tec1Δ (PC6102), and the tec1Δ ssa1Δ double mutants (PC7819) grown on YEPD plates for 16 h. Scale bar, 10 µm. (F) Left: Microscopic examination of wild-type cells (WT, PC538), rsp5-1 (PC7543), tec1Δ (PC6102), and the tec1Δ rsp5-1 double mutants (PC7826) grown on YEPD plates for 16 h. Scale bar, 10 µm. Right: Quantification of cell length-to-width ratio of same cells, from three biological replicates (n = 3); >200 cells were examined in each replicate. Data were analyzed by one-way ANOVA, and P values (asterisk < 0.01) were calculated using Tukey’s multiple comparison test. (G) Left: Wild-type cells (WT, PC3288) and the rsp5-1 mutant (PC3290) expressing GFP-Cdc42p (Cdc42, PC6454) or GFP-Cdc42pQ61L (Q61L, PC7458) were examined by microscopy. Scale bar, 5 µm. Right: Quantification of cell length of same cells, n > 25. Data were analyzed by one-way ANOVA, and P values (asterisk < 0.01) were calculated using Tukey’s multiple comparison test.

Role of the HSP40 Ydj1p, the HSP70 Ssa1p, and Rsp5p in regulating fMAPK activity. (A) Serial dilutions of wild-type cells (WT, PC538), ydj1Δ (PC7616), and ssa1Δ (PC7621) mutants grown on YEPD plates for 2 d. (B) Fluorescence microscopy of wild-type cells (WT, PC986) and the ydj1Δ mutant (PC7657) in the S288c background expressing GFP-Cdc42p (PC6454). Scale bar, 5 µm. (C) Fluorescence microscopy of wild-type cells (WT, PC6016) and the ssa1Δ mutant (PC7700) expressing GFP-Cdc42p (PC6454). Scale bar, 5 µm. (D) Microscopic examination of wild-type cells (WT, PC538), ydj1Δ (PC7616), tec1Δ (PC6102), and the tec1Δ ydj1Δ double mutants (PC7820) grown on YEPD plates for 16 h. Scale bar, 10 µm. (E) Microscopic examination of wild-type cells (WT, PC538), ssa1Δ (PC7621), tec1Δ (PC6102), and the tec1Δ ssa1Δ double mutants (PC7819) grown on YEPD plates for 16 h. Scale bar, 10 µm. (F) Left: Microscopic examination of wild-type cells (WT, PC538), rsp5-1 (PC7543), tec1Δ (PC6102), and the tec1Δ rsp5-1 double mutants (PC7826) grown on YEPD plates for 16 h. Scale bar, 10 µm. Right: Quantification of cell length-to-width ratio of same cells, from three biological replicates (n = 3); >200 cells were examined in each replicate. Data were analyzed by one-way ANOVA, and P values (asterisk < 0.01) were calculated using Tukey’s multiple comparison test. (G) Left: Wild-type cells (WT, PC3288) and the rsp5-1 mutant (PC3290) expressing GFP-Cdc42p (Cdc42, PC6454) or GFP-Cdc42pQ61L (Q61L, PC7458) were examined by microscopy. Scale bar, 5 µm. Right: Quantification of cell length of same cells, n > 25. Data were analyzed by one-way ANOVA, and P values (asterisk < 0.01) were calculated using Tukey’s multiple comparison test.

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