Figure 2.

Proper Cdc42 function is required for Bgs1 recruitment to the medial ring during cytokinesis. (a). Sum projections showing GFP-Bgs1 and Rlc1-tdTomato localization at permissive (25°C) and non-permissive (36°C) temperatures in cdc42⁺ and cdc42-1625 cells, and respective DIC images. Middle panel shows middle of z-section with Rlc1-tdTomato signal to indicate how we classified non-constricting rings (two distinct dots). The white arrows point to GFP-Bgs1 localization at division site in cells with non-constricting rings. (b). Sum projections of cdc42⁺ and cdc42-1625 cells with magenta box showing 330 pixel² box used to measure GFP-Bgs1 signal at the division site. ⁽c). Fold change in GFP-Bgs1 fluorescence intensity, in cells with non-constricting rings in the indicated strains; in comparison to the intensity in cdc42⁺ cells at 25°C, n = 3 replicate experiments, with >32 cells for each strain in each experiment. (d). Western Blot showing global GFP-Bgs1 levels at 25°C and 36°C. (e). Quantification of the percentage of cells with rings that display Bgs1 localization, n = 2 replicate experiments, with ≥100 rings analysed per strain. (f). Quantification of the percentage of constricting rings observed among cells with an actomyosin ring in the mentioned strains, at 25°C and 36°C, number of rings analysed(n) ≥100 cells for each strain. The red lines on graph represent mean, error bars represent standard deviation; Statistics is performed with ANOVA, followed by Tukey posthoc analysis, *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001; n.s., not statistically significant; Scale bars- 5 µm