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Image Search Results
Journal: microPublication Biology
Article Title: Characterization of temperature-sensitive alleles of the septation initiation network protein Mob1 in Schizosaccharomyces pombe
doi: 10.17912/micropub.biology.001595
Figure Lengend Snippet: (A) The mutations encoded by each mob1 allele are listed. (B) The indicated strains were grown in liquid YE media at 25°C until they reached mid-log phase and then adjusted to the same cell concentrations measured by optical density (Moreno et al., 1991). Next, 10-fold serial dilutions were made and 2.5 µL of each was spotted on YE agar plates and incubated at the indicated temperatures for 2-3 days prior to imaging. The spot assays were done twice and a representative is shown. (C) The indicated strains were grown in liquid YE media at 25˚C. Samples were collected before and again after growing the cells for an additional 3.5 hours at 36˚C. The cells were then fixed and stained with DAPI and methylene blue. Representative images are shown. The experiment was performed in duplicate. Scale bar, 5 µm. (D) The number of nuclei per cell (top), and the percentage of septated cells (middle) and lysed cells (bottom) were quantified at 36°C from the same experiments as in C. N≥200 cells of each genotype. (E) Ribbon diagram of a structural model of S. pombe Mob1 using AlphaFold3 (Abramson et al., 2024). The positions of the N- and C-termini and the positions of the mutated residues in the mob1 alleles are indicated. (F) Schematic of sid2 gene product (drawn to scale). Numbers indicate amino acid position (top). Ribbon diagram of a structural model of S. pombe Mob1 bound to the Sid2 regulatory region and kinase domain using AF3. Mob1 is in magenta, the Sid2 regulatory region is in green, and the Sid2 kinase domain is in cyan (bottom).
Article Snippet:
Techniques: Comparison, Incubation, Imaging, Staining