Results: 4

1.
Figure 1

Figure 1. From: The Saccharomyces SUN gene, UTH1, is Involved in Cell Wall Biogenesis.

UTH1 is involved in the yeast cell's response to different stresses. 5 μl aliquots of 10-fold serial dilutions of wildtype and Δuth1 mutant cells in the W303-1A strain background were plated onto the designated media and cultured at either 30°C or 39°C for 2, 3 or 5 days, depending upon the particular plate. All spot assays were repeated at least three times and a representative experiment is shown.

James J. Ritch, et al. FEMS Yeast Res. ;10(2):168-176.
2.
Figure 3

Figure 3. From: The Saccharomyces SUN gene, UTH1, is Involved in Cell Wall Biogenesis.

Deleting UTH1 decreases the rate of spheroplast formation of yeast cells. Cells of the indicated genotypes were cultured overnight in rich YPD media at 30°C, harvested and washed three times with deionized water, and then resuspended to an OD660 of 0.5 in TE buffer, pH 7.5 (50 mM Tris/HCl, 5 mM EDTA). Zymolyase (5 U/μl; Zymo Research, Orange, CA) was then added to the cells to a final concentration of 12 μg/ml. Cell suspensions were incubated at 23°C and their optical density (OD660) was recorded at four-minute intervals for the indicated time period. Assays were done in triplicate and mean values are shown along with the standard deviation.

James J. Ritch, et al. FEMS Yeast Res. ;10(2):168-176.
3.

Figure 2. From: The Saccharomyces SUN gene, UTH1, is Involved in Cell Wall Biogenesis.

UTH1 is involved in maintaining the integrity of the yeast cell wall. (A) Deleting UTH1 enhances cell growth on media supplemented with calcofluor white (CFW) or sodium dodecyl sulfate (SDS). 5 μl aliquots of 10-fold serial dilutions of wildtype and Δuth1 mutant cells in the W303-1A strain background were plated onto the designated media and cultured at 30°C for 2 or 4 days, depending upon the particular plate. YPDS denotes YPD plates supplemented with 1 M sorbitol respectively. (B) Overexpression of PKC1 enhances cell wall integrity mimicking a deletion of UTH1. 5 μl aliquots of 10-fold serial dilutions of wildtype and Δuth1 mutant cells transformed with plasmid pFR22 to overexpress PKC1 were plated onto the designated selective SD plates and cultured at 30°C for 2 or 5 days. SD-S denotes SD plates supplemented with 1 M sorbitol. All spot assays were repeated at least three times and a representative experiment is shown.

James J. Ritch, et al. FEMS Yeast Res. ;10(2):168-176.
4.

Figure 4. From: The Saccharomyces SUN gene, UTH1, is Involved in Cell Wall Biogenesis.

Deleting UTH1 enhances the cell's response to diverse stresses by strengthening its cell wall. (A) Osmotic stabilization with 1M sorbitol phenocopies a deletion of UTH1. 5 μl aliquots of 10-fold serial dilutions of wildtype and Δuth1 mutant cells were plated onto the designated media and cultured at either 30°C or 39°C for 2, 3 or 5 days. YPDS and SD-S denotes YPD and SD plates supplemented with 1 M sorbitol respectively. To overexpress human BAX in our strains, we transformed plasmid pCM189/Bax into our cells and plated them on selective SD media with or without doxycycline supplementation as previously described (Camougrand et al., 2003). (B) Overexpression of PKC1 enhances cell growth at elevated temperatures mimicking a deletion of UTH1. 5 μl aliquots of 10-fold serial dilutions of wildtype and Δuth1 mutant cells transformed with plasmid pFR22 to overexpress PKC1 were plated onto the designated selective SD plates and cultured at 30°C or 38°C for 2 or 5 days. All spot assays were repeated at least three times and a representative experiment is shown.

James J. Ritch, et al. FEMS Yeast Res. ;10(2):168-176.

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