FIG. 2.

Ndc10, Ctf3, and Cse4 coimmunoprecipitation with CEN DNA is not dependent on Rsc2. Anti-Myc or anti-HA ChIP assays were performed with chromatin extracts of wild-type (RSC2) or rsc2Δ mutant cells expressing either Ndc10-Myc (YVM499 and YKB121), Ctf3-Myc (YVM 218 and YKB127), or Cse4-HA (YVM1141 and YKB124), as well as with cells expressing no tag (YPH499). Cells were grown to mid-log phase in YPD medium at 25°C, and then the culture was shifted to 37°C for 3 h before chromatin was isolated for IP. Multiplex PCR was performed to amplify the centromeric DNA of CEN1, CEN3, and a non-CEN locus, PGK1, for both whole-cell extracts (WCE) and immunoprecipitates.