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Fig. 6. Association of Rec8 with chromosomes and its retention at centromeres in meiosis II are not affected in the absence of CTF8. Diploid wild-type (K10003) (A,C) and ctf8{Delta} (K10572) (B,D) strains expressing Rec8-HA3 and Pds1-Myc18 (securin) were analysed in a meiotic time-course experiment by chromosome spreading (A,B) and in situ immunofluorescence (C,D). Chromosome spreads were stained for DNA and with antibodies against the HA tag. Pachytene-stage chromosome spreads of wild-type (A) and ctf8{Delta} (B) cells showing Rec8 (red) and DNA (blue) in the upper panels and Rec8 alone (white) in the lower panels. Fixed cells were stained for DNA and with antibodies against tubulin, the HA and Myc epitope, and analysed by fluorescence microscopy. Wild-type (C) and ctf8{Delta} (D) metaphase II cells showing tubulin (red) and DNA (blue) in the upper panel, Rec8 (white) in the middle panel, and securin (white) in the lower panel. 100 cells were scored for each strain in this experiment.