Fig. 5. Localisation of flp 1p-GFP in SIN mutants. Spores were inoculated into
minimal medium containing adenine and leucine, and allowed to germinate
(except D1-4). The localisation of flp 1p-GFP was examined in multinucleate
cells. (A) cdc7 null allele. (1) Interphase, binucleate cell: note
that flp 1p-GFP is present in the nucleolus and on the spindle pole body. (2)
Cell undergoing a second mitosis: note the presence of a single medial ring
and staining along the spindle. (B) cdc14 null allele. (1) Binucleate
cell, early in second mitosis: note the presence of the medial ring, and the
spindle staining. (2) Multinucleate interphase cell: note that flp 1p-GFP is
present in the nucleolus and on the spindle pole body. (C) spg1 null
allele. (1) Mitotic cell: note the presence of two medial rings. (2) Mitotic
cell: note the presence of staining along the short spindle and on the spindle
pole bodies. (3) Multinucleate interphase cell: note that flp 1p-GFP is
present in the nucleolus and on the spindle pole body. (Inset) Binucleate cell
early in the second mitosis: note the spindle staining. (D) cdc16
null allele, germinating cells were fixed and processed for indirect
immunofluorescence. flp 1p-GFP was detected with rabbit antiserum against GFP
(2,4); DNA was detected with DAPI (1,3). Images 1-4 are type I cells. Note
that there is no discrete nucleolar or spindle pole body staining of flp
1p-GFP. Spotty aggregates were sometimes observed (arrows). Image 5 is a type
II cell, nucleolar and spindle pole body staining of flp 1p-GFP is observed.
(E) sid2-1 cdc16::ura4+ flp 1-GFPc cells were
grown at 25°C, and the GFP fluorescence was observed in living cells. The
aggregate observed in cell 2 is shown by the arrow. Images 1 and 3 are
phase-contrast views of the cells shown in images 2 and 4, respectively.
