Figure 1.

Formation of Neocentromeres on Chromosome Z

(A) A strategy to isolate surviving cells after removal of endogenous centromere of chromosome Z with positive (Zeocin resistance) and negative (FIAU resistance) selections. Surviving cells are expected to have chromosome Z with a neocentromere or fusion chromosome Z with another chromosome.

(B) Southern hybridization analysis to confirm that endogenous centromere sequence is removed. Probe information (probe #1–5) for Southern analysis to confirm removal of endogenous centromere is also shown.

(C) Frequency of isolation of surviving cells without endogenous centromere sequence. We independently performed four trials.

(D) Classification of surviving clones based on cytological experiments using anti-CENP-T (CT) antibodies as a centromere marker. Among 136 clones, ten clones have fusion chromosome Z with another chromosome. We classified the centromere position defined by CENP-T staining as five types (type I–V).

(E) Immuno-FISH images of various types of chromosomes. Red shows signals from the satellite sequence on q-arm of chromosome Z. Centromeres (green) are visualized by anti-CENP-T antibodies.

See also Figure S1.