Fig. 2. Dis2.NEGFP associates with centromeres. dis2.NEGFP pcp1.RFPC (IH2731; A,B) and dis2.NEGFP cnp1.Cherry (IH5283; C,D) cells were processed as for Fig. 1 with the exception that DMSO or DMSO containing CBZ at a final concentration of 25 µg/ml were included in the culture medium for imaging in A and B, respectively. (A,B) The Dis2.NEGFP signal does not colocalise with the Pcp1.RFPC signal of interphase cells. (C,D) The Dis2.NEGFP signal colocalises with the Cnp1.Cherry signal of interphase (C) and mitotic (D) cells. Inset in D shows enlargement of the nucleus marked with the asterisk. (E) For ChIP analysis whole cell extracts (wce) were prepared from the indicated strains for precipitation with magnetic beads to which either no antibody (–) or a rabbit antibody that recognised the Pk epitope (+) had been covalently attached. There was a clear enrichment of the signal arising from PCR with primers to the central but no other sequences in immunoprecipitates from dis2.NPk (lane 5), but not when antibodies were not conjugated to the beads (lane 4) or the strain did not contain the tagged dis2⁺ gene (lane 2). Bars, 5 µm.