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. 2010 Sep 6;190(5):793–805. doi: 10.1083/jcb.201002055

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© 2010 Ray et al.

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Figure 5. — The MOR pathway mutant orb6-25 rescues growth and cell division defects of SIN mutants. (A) Cells of the indicated genotypes were grown at 25°C and their growth was tested at the indicated temperatures by spotting 10-fold serial dilutions on YE agar plates. (B) Cells of the indicated genotypes were shifted to 36°C for 4 h, methanol fixed, and stained with Calcofluor White (CW) to score for septum formation. Representative cdc7-24 (montage) and cdc7-24 orb6-25 cells show the septation status in respective cells and arrows mark the septum. Dashed dividing lines separate the individual images in the montage. Arrows mark the septum in the cdc7-24 orb6-25 cells. The septation index (percentage of cells with septa) for all the genotypes indicated is shown in the histogram plot. At least 100 cells were scored for each strain. Error bars denote the SD for three separate experiments. (C) Time-lapse images of the indicated strains were acquired using a spinning disc confocal microscope (TE 2000-E2; Nikon). Cells were grown at 25°C and then placed on the microscope stage that was maintained at 36°C during the entire time of imaging. Arrowheads indicate completion of ring constriction in representative cells. (D) Cells of the indicated genotypes were grown as in B and the proportion of cells with ectopic septa (either septa present in mononucleate cells, or multiple septa) was scored. At least 100 cells were scored for each genotype. The experiment was done in triplicate and error bars denote the SD values. Arrows in the montage image point to the ectopic septa found in representative cells. Dashed dividing lines separate the individual images in the montage.