Figure 9.
RacC is required to maintain cortical integrity. (A) WT, racC−, and GFP-RacC/racC− (rescue) cells grown for 60 h in shaken suspension were fixed and stained with DAPI to visualize the nuclei. (B) Quantification of nuclei in cells. n, number of cells analyzed. (C) Top: Trajectories of randomly migrating cells (n = 65 for WT and 69 for racC−). Bottom: Summary of motility parameters. (D and E) Velocity and Euclidean distance of cells are shown in C. At least 15 cells were quantified per experiment (each experiment shown in a different color); mean ± SEM. (F) Top: Trajectories of cells migrating under 2% agarose along folic acid gradient (n = 76 for WT and 75 for racC−). Bottom: Summary of chemotaxis parameters. (G–I) Speed, FMI, and directness of cells are shown in F. At least 20 cells were quantified per experiment (each experiment shown in a different color); mean ± SEM. (J) Localization of GFP-ABD in WT and racC− cells. (K) Quantification of the cortex-to-cytoplasm fluorescent intensity ratios of GFP-ABD. (L) WT and racC− cells were fixed and stained with Alexa Fluor 555-labeled phalloidin. (M) Quantification of the fluorescence intensity of cortical phalloidin. (N) Projection length (Lp) of WT and racC− cells determined by micropipette aspiration using a constant pressure of 500 Pa for 5 min. (O) Quantitative analysis of the projection lengths of probed cells. For C and F, data were from three independent experiments (the average of each biological replicate was used to calculate the mean and SEM). For K, M, and O, data were from three independent experiments; the scatter plots show data points with mean ± SEM; n, number of cells analyzed. Scale bars, 5 μm.

RacC is required to maintain cortical integrity. (A) WT, racC−, and GFP-RacC/racC− (rescue) cells grown for 60 h in shaken suspension were fixed and stained with DAPI to visualize the nuclei. (B) Quantification of nuclei in cells. n, number of cells analyzed. (C) Top: Trajectories of randomly migrating cells (n = 65 for WT and 69 for racC−). Bottom: Summary of motility parameters. (D and E) Velocity and Euclidean distance of cells are shown in C. At least 15 cells were quantified per experiment (each experiment shown in a different color); mean ± SEM. (F) Top: Trajectories of cells migrating under 2% agarose along folic acid gradient (n = 76 for WT and 75 for racC−). Bottom: Summary of chemotaxis parameters. (G–I) Speed, FMI, and directness of cells are shown in F. At least 20 cells were quantified per experiment (each experiment shown in a different color); mean ± SEM. (J) Localization of GFP-ABD in WT and racC− cells. (K) Quantification of the cortex-to-cytoplasm fluorescent intensity ratios of GFP-ABD. (L) WT and racC− cells were fixed and stained with Alexa Fluor 555-labeled phalloidin. (M) Quantification of the fluorescence intensity of cortical phalloidin. (N) Projection length (Lp) of WT and racC− cells determined by micropipette aspiration using a constant pressure of 500 Pa for 5 min. (O) Quantitative analysis of the projection lengths of probed cells. For C and F, data were from three independent experiments (the average of each biological replicate was used to calculate the mean and SEM). For K, M, and O, data were from three independent experiments; the scatter plots show data points with mean ± SEM; n, number of cells analyzed. Scale bars, 5 μm.

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