Figure S2.
GTP-Cdc42 localization and cell size in cells treated with nocodazole or latrunculin.(A) Representative time-lapse image showing the localization of Gic2-PBD during mitosis in a wild-type cell treated with nocodazole. Numbers indicate time in minutes from the initial polarization of Cdc42 in G1. Cells were synchronized in G1 with α factor and released into medium containing nocodazole. Scale bar: 5 µm. (B) Percent of wild-type cells without nocodazole and with nocodazole with Gic2-PBD bud concentration over time (n = 100 cells from three experiments for each background). (C) Gic2-PBD-RFP fluorescence intensity in cells treated with nocodazole for two hours. Average intensity from each cell compartment was normalized to the total intensity in the cell (n = 50 cells from three experiments; average ± SEM). Asterisks indicate statistically significant differences (Mann-Whitney t test; P < 0.0001). (D) Cell volume of buds and mothers of cells treated with nocodazole (n = 30 cells from three experiments for each time point indicated; average ± SEM). (E) Representative time-lapse image showing the localization of Gic2-PBD in a cell treated with LatB. Numbers indicate time in minutes from the initial polarization of Cdc42 in G1. Scale bar, 5 µm. For E–I, cells were synchronized in G1 with α factor, released into the cell cycle and arrested in M phase by CDC20 depletion. (F) Growth rate of buds versus mothers with and without LatB treatment (n = 30 cells from three experiments; average ± SEM). The growth rate was measured between 0 and 180 min, where time 0 is the time of LatB addition. Asterisks indicate statistically significant differences (****, P < 0.0001, Mann-Whitney test). (G) Cell volume of mothers and buds with and without LatB treatment (n = 30 cells for every time point indicated from three experiments; average ± SEM). Asterisks indicate statistically significant differences (****, P < 0.0001, Mann-Whitney t test). (H) Average cell volume after 3 h of LatB treatment (n = 30 from three experiments; average ± SEM). (I) Percentage of cells with spindle migration to the bud with or without LatB treatment (n = 100 cells for each category from three experiments).

GTP-Cdc42 localization and cell size in cells treated with nocodazole or latrunculin. (A) Representative time-lapse image showing the localization of Gic2-PBD during mitosis in a wild-type cell treated with nocodazole. Numbers indicate time in minutes from the initial polarization of Cdc42 in G1. Cells were synchronized in G1 with α factor and released into medium containing nocodazole. Scale bar: 5 µm. (B) Percent of wild-type cells without nocodazole and with nocodazole with Gic2-PBD bud concentration over time (n = 100 cells from three experiments for each background). (C) Gic2-PBD-RFP fluorescence intensity in cells treated with nocodazole for two hours. Average intensity from each cell compartment was normalized to the total intensity in the cell (n = 50 cells from three experiments; average ± SEM). Asterisks indicate statistically significant differences (Mann-Whitney t test; P < 0.0001). (D) Cell volume of buds and mothers of cells treated with nocodazole (n = 30 cells from three experiments for each time point indicated; average ± SEM). (E) Representative time-lapse image showing the localization of Gic2-PBD in a cell treated with LatB. Numbers indicate time in minutes from the initial polarization of Cdc42 in G1. Scale bar, 5 µm. For E–I, cells were synchronized in G1 with α factor, released into the cell cycle and arrested in M phase by CDC20 depletion. (F) Growth rate of buds versus mothers with and without LatB treatment (n = 30 cells from three experiments; average ± SEM). The growth rate was measured between 0 and 180 min, where time 0 is the time of LatB addition. Asterisks indicate statistically significant differences (****, P < 0.0001, Mann-Whitney test). (G) Cell volume of mothers and buds with and without LatB treatment (n = 30 cells for every time point indicated from three experiments; average ± SEM). Asterisks indicate statistically significant differences (****, P < 0.0001, Mann-Whitney t test). (H) Average cell volume after 3 h of LatB treatment (n = 30 from three experiments; average ± SEM). (I) Percentage of cells with spindle migration to the bud with or without LatB treatment (n = 100 cells for each category from three experiments).

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