neo1R neocentromere clusters with endogenous centromeres at the SPB during interphase
(A) Diagram represents strains with cen1 or lacking cen1 but carrying neo1R neocentromere (cen1Δ neo1R). Red line indicates position of neo1R DNA FISH probe (ChrI: 5,513,871–5,530,124).
(B and D) Representative images of neo1R DNA FISH (red; probe as indicated in A), SPB location (green; anti-Cdc11; B) or centromere clusters (green; anti-CENPCnp1; D), and DNA staining (blue, DAPI) in WT cen1 (B) and cen1Δ neo1R cells. Images were scaled as in Figure 1. Scale bars, 5 μm.
(C and E) 3D distances between neo1R DNA and SPBs (Cdc11; C) or centromere clusters (CENP-ACnp1; E).
Percentage of interphase cells (n, number analyzed) in each distance category, classified as in Figure 1. AV, average distance. ∗∗∗p < 0.0001 (Mann-Whitney U test) (see also Figure S1).