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Fig. 3. The mating-type region is localized close to the SPB at the nuclear periphery in the wild-type strains and is delocalized in strains with cis- and trans-acting silencing mutations. (A) Confocal microscopy images of wild-type and mutant cells. Shown in the different panels are: the NM labelled with DsRed (red, first column), the SPB labelled with CFP (blue, second column), the mating-type region or (in the bottom row) the cut3 locus, labelled with lacR-GFP (green, third column), and merged images (fourth column). Bars, 2 µm. Histograms with the distributions of observed distances between the mating-type region (or, in the bottom row, the cut3 locus) and the SPB are shown to the right of the panels. The distribution of the observed distances between the GFP and CFP signals were binned in groups spanning 0.2 µm. The strains analysed were PJ644 (w.t. ade6-M210), PJ654 (w.t. ade6-DN/N), PJ579 (IR-L/IR-R), PJ577 (h^K repressed) and PJ586 (h^K derepressed), PJ653 (clr4), PJ668 (swi6), PJ651 (dcr1) and PJ575 (cut3::lacO). (B) The nucleus divided into three zones of equal surface area. Zone I corresponded to a distance of 0-0.22 µm, zone II to a distance of 0.23-0.51 µm and zone III to a distance of 0.52-1.20 µm from the nuclear periphery. (C) Distribution of mat2/3 localization into zone I, zone II or zone III for each strain. The significance of the differences between each mutant strain and the PJ644 wild type was tested statistically (Mann-Whitney rank sum test, P<0.05). no, no significant difference; yes, a significant difference; n.a., not applicable. (D) The distribution of cells having the mat2/3 region localized into zone I, zone II or zone III plotted as pie charts. Black, zone I; light grey, zone II; dark grey, zone III.

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