Additional file 1 of Sphingolipids are required for exocyst polarity and exocytic secretion in Saccharomyces cerevisiae
Creators: name: Guo, Qingguo
name: Zhang, Tianrui
name: Meng, Na
name: Duan, Yuran
name: Meng, Yuan
name: Sun, Dong
name: Liu, Ying
name: Luo, Guangzuo
Publisher: name: figshare
Year: 2020
Subjects: subject: Biophysics
subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Physiology
subject: Biotechnology
subject: Immunology
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Cancer
subject: Hematology
subject: Infectious Diseases
subject: FOS: Health sciences

description: Additional file 1: Figure S1. The amount of the GFP tagged exocyst subunits did not differ significantly change in lcb1-100 mutant and cells treated with myriocin. (A) The yeast strains harboring SEC3-GFP, SEC5-GFP, SEC8-GFP or EXO70-GFP on the chromosome were grown to early log phase in YPD liquid medium overnight at 25°C. The cells were then shifted from 25℃ to 39℃ for 60 min. Total protein was extracted and Western blots were performed using rabbit anti-GFP antiby (1:1000 dilution). Adh1 was used as a loading control (rabbit anti-Adh1 antibody). (B) Wild-type cells harboring SEC3-GFP, SEC5-GFP, SEC8-GFP or EXO70-GFP on the chromosome were grown to early log phase in YPD liquid medium overnight at 25°C. The cells were then treated with or without 10μg/ml myriocin for 60 min. Total protein was extracted and Western blots were performed using rabbit anti-GFP antiby (1:1000 dilution). Adh1 was used as a loading control (rabbit anti-Adh1 antibody). Figure S2. Cell viability of wild type and lcb1-100 mutant cells at a restrictive temperature. (A) lcb1-100 mutant is temperature sensitive. Wild type and lcb1-100 mutant cells were grown to log phase in YPD at 25°C, and equal densities of wild-type or lcb1-100 mutant cells were spotted in 10-fold serial dilutions from left to right on YPD plate, and then incubated at different temperatures for 3 days. (B) Viability of lcb1-100 mutant cells at restrictive temperature. Wild-type and lcb1-100 mutant cells were grown to early log phase in YPD liquid medium overnight at 25°C and then shift to 39°C for 60 min. The cells were stained with 4% Loeffler’s methylene blue solution for 10min at room temperature. Cells were washed with PBS buffer and examined by microscopy. Cells stained with blue color are not viable. (C) Statistical analysis of cell viability in wild type and lcb1-100 mutant cells in (B). n=3. Three independent experiments were performed. *, p&lt;0.01. Figure S3. Cell viability of wild type cells treated with or without myriocin. (A) Wild-type and lcb1-100 mutant cells were grown to early log phase in YPD liquid medium overnight at 25°C. Myriocin was added into the medium to a final concentration of 10μg/ml, and the cells were incubated at 25°C for 60 min. The cells were collected and stained with 4% Loeffler’s methylene blue solution for 10 min at room temperature. Cells were washed with PBS buffer and examined by microscopy. Cells stained with blue color are not viable. (B) Statistical analysis of viability of wild type cells treated with myriocin in (A). n=3. Three independent experiments were performed. *, p&lt;0.01. Table S1. Yeast strains used in this study. Table S2. Plasmids used in this study.