Figure 2. A probe for the visualization of Ras-GTP. (A) Schematic representation of MAP2K Byr2 (top) and probes to visualize Ras-GTP (RasActGFP) in S. pombe (S.p.; middle) and S. cerevisiae (S.c.; bottom). Promoters and terminators used for gene expression are indicated. (B and C) Localization of RasActmCherry and GFP-Ras1 (B), and RasActGFP (C), during vegetative growth of S. pombe cells. Note that the nuclear and dotted (mitochondrial) localization of RasActGFP are nonspecific, as they do not depend on ras1. (D) Cortical tip profiles of RasActGFP fluorescence in strains as in C; n = 25. Thick line, mean; shaded area, SD. (E) Localization of RasActGFP during vegetative growth of S. c. cells. (F) Mean total RasActGFP cortical fluorescence in strains as in E; n = 25; ***, 3.9 × 10−13 ≤ P ≤ 3.7 × 10−5. Error bars, SD. Bars, 2 µm.
Figure 2.

A probe for the visualization of Ras-GTP. (A) Schematic representation of MAP2K Byr2 (top) and probes to visualize Ras-GTP (RasActGFP) in S. pombe (S.p.; middle) and S. cerevisiae (S.c.; bottom). Promoters and terminators used for gene expression are indicated. (B and C) Localization of RasActmCherry and GFP-Ras1 (B), and RasActGFP (C), during vegetative growth of S. pombe cells. Note that the nuclear and dotted (mitochondrial) localization of RasActGFP are nonspecific, as they do not depend on ras1. (D) Cortical tip profiles of RasActGFP fluorescence in strains as in C; n = 25. Thick line, mean; shaded area, SD. (E) Localization of RasActGFP during vegetative growth of S. c. cells. (F) Mean total RasActGFP cortical fluorescence in strains as in E; n = 25; ***, 3.9 × 10−13 ≤ P ≤ 3.7 × 10−5. Error bars, SD. Bars, 2 µm.

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