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. 2008 Jul 11;4(7):e1000120. doi: 10.1371/journal.pgen.1000120

Figure 7. Overexpression of RFA1 causes chromosomal segregation and spindle defects.

Figure 7

(A) The G2/M delay phenotype upon overexpression of RFA1 is apparent in flow cytometry histograms of the empty vector control strain and the PGAL1-RFA1 strain. (B) Quantitation of cell microscopy results following nuclear DNA staining indicates that PGAL1-SPC97 and PGAL1-RFA1 strains exhibit considerably higher percentages of large-budded cells with undivided nuclei than the empty vector control strain. (C) Log-phase cultures of the wild type control strain and cells carrying PGAL1-SPC97 or PGAL1-RFA1 were fixed in formaldehyde and stained to visualize DNA (by DAPI) and microtubules (by immunofluorescence). The cells carrying the empty vector correctly showed a long anaphase spindle, with nuclei successfully segregated into two cell bodies. Overexpression of either SPC97 or RFA1 resulted in a failure of chromosome segregation; the spindle morphology of PGAL1-RFA1 cells is distinct from that of PGAL1-SPC97 cells, with shorter mitotic spindles poorly aligned with the division axis.