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. 2019 May 3;8:e46003. doi: 10.7554/eLife.46003

Figure 5. Redistribution of Pom1 clusters to the lateral cortex disrupts Cdr2 nodes during glucose restriction.

(A) Localization of Pom1-mNG in cells grown in either high (2%) or low (0.03%) glucose media. Images were collected using TIRF microscopy. Scale bar, 5 µm. (B) Comparison of the binding frequency of Pom1 clusters at the lateral cell cortex in cells grown under high and low glucose (***p=0.0003, n = 10 cells, 32–113 traces/cell). Statistical significance was tested using a Student’s T-test. (C) Comparison of the binding dwell time of Pom1 clusters at the lateral cell cortex in cells grown under high and low glucose (n.s., p=0.6833, n = 10 cells, 32–113 traces/cell). Statistical significance was tested using a Student’s T-test. (D) Localization of Cdr2-mNG in wild-type or pom1∆ cells grown in either normal (2%) or low (0.03%) glucose media. Images were collected using TIRF microscopy. Scale bar, 1 µm. (E) Comparison of the fluorescence intensity of individual Cdr2-mNG nodes in wild-type or pom1∆ cells grown in either normal (2%) or low (0.03%) glucose media. Low glucose induces partial node disassembly in wild-type cells (****p<0.0001, n = 92–169 nodes from >5 cells) but not in pom1∆ cells (n.s., p=0.4015, n = 113–156 nodes from >5 cells). Measurements were taken from Airyscan Super-Resolution confocal micrographs. (F) Quantification and comparison of the total number of Cdr2-mNG nodes visible in TIRF micrographs of wild-type or pom1∆ cells grown in either normal (2%) or low (0.03%) glucose media. There is no significant difference in any condition (p>0.05, n = 28–33 cells). Statistical significance was tested using a one-way ANOVA.

Figure 5.

Figure 5—figure supplement 1. Redistribution of Pom1 clusters to the lateral cortex disrupts Cdr2 nodes during glucose restriction, supporting data.

Figure 5—figure supplement 1.

(A) Analysis of Pom1 and Cdr2 redistribution by spinning-disc confocal microscopy, as in Kelkar and Martin (2015). Images are confocal micrographs of cells co-expressing Pom1-tdTomato and Cdr2-mNG in high (2%) or low (0.03%) glucose media. Scale bar, 5 µm. (B) Cortical fluorescence intensity of Cdr2 and Pom1 was measured using line scans drawn around the circumference of the representative cell marked with an arrowhead in the 2% glucose portion of panel A. (C) Cortical fluorescence intensity of Cdr2 and Pom1 was measured using line scans drawn around the circumference of the representative cell marked with an arrowhead in the 0.03% glucose portion of panel A.