Analysis of the cell cycle arrest in
ntf2–2ts mutant cells. (A) DNA content in
NTF2 mutants. NTF2,
ntf2–1ts, ntf2–2ts, MAD2Δ,
ntf2–2ts MAD2Δ, and ntf2–1ts MAD2Δ
cells were grown overnight at 25°C, were diluted to 2 ×
106 cells/ml in YEPD, and were shifted to 37°C, and
samples were taken at 0, 4, 6, and 8 h. The DNA content of
individual NTF2, ntf2–1ts, and
ntf2–2ts cells was determined by flow cytometry. (B)
Spindle pole body localization in NTF2 mutants.
NTF2, ntf2–1ts and
ntf2–2ts cells were transformed with a plasmid encoding
Nuf2-GFP (Kahana et al., 1995) to visualize spindle pole
bodies. Transformed cells were grown to log phase at 25°C and then
shifted to 37°C for 3 h. GFP-labeled spindle pole bodies were
visualized by direct fluorescent microscopy.