Fig. 6. Nuf2p and Mcm21p are located close to the CEN5. (A) Localization of Mcm21p-9Myc, Nuf2p-9Myc and Spc98p-9Myc relative to CEN5 DNA. Synchronized cells with GFP-labeled CEN5 DNA carrying MCM21-9Myc, NUF2-9Myc or SPC98-9Myc were fixed and analysed by direct and indirect immunofluorescence for CEN5 DNA (green), Myc-tagged protein (red) and tubulin (blue). Note that the overlap between the green and red signals appears in yellow. Shown are cells in G₁ phase (monopolar spindle), G₂/S phase (short spindle) and anaphase (long spindle) of the cell cycle. Bar: 5 µm. (B) Quantification of (A). Cells (n = 100) in G₁, G₂/S phase or anaphase of the cell cycle were analysed. Signals were counted as ‘co-localized’ when the CEN5 signal (green) was <0.4 µm from the red anti-HA dot.