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Fig. 3. (A) Flow cytometry of cdc37-184, cdc37-J, cdc37+ ED1022 and cdc10-129 to determine the DNA content of cells. Strains were cultured in YE at 28 and 36°C over an 8 hour time course. Samples of cells were taken every 2 hours and processed for flow cytometry. (B) Frequency of phenotypes observed by DAPI staining of cdc37-184, cdc37-681 and cdc37+ cells. Samples of cells were taken every 2 hours, fixed in formaldehyde and stained with DAPI. Phenotypes 1,2 and 3 are shown in C. Phenotype 4 is a cell with a single nucleus and a septum, phenotype 5 is a cell with a septum cutting through a single nucleus and phenotype 6 is a cell with multiple septa. (C) Cellular phenotypes 1, 2 and 3 observed with DAPI staining of cdc37+ and mutants at both 28 and 36°C. (D,E,F) Immunofluorescence of microtubules, using the TAT1 antibody, of cdc37+ interphase microtubules (D), and arrested cdc37-184 (E) and cdc37-J (F) cells. Strains were cultured at 28 and 36°C for 8 hours. (G) Immunofluorescence of cdc37-184 and cdc37+ ED1022 cells with the anti-S. pombe Cdc37 antibody. Samples of cells were processed for immunofluorescence with anti-S. pombe Cdc37 antibody or depleted antibody (see Materials and Methods). Bars, 10 µm.





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