Figure 5.
Dam1p is at the kinetochore. (A) IF of chromosome spreads from cells containing Ndc10pHA and Dam1p-myc. Blue shows DAPI (DNA), green shows Ndc10pHA detected with anti-HA mAb plus FITC anti-mouse secondary Ab, and red shows Dam1pMyc detected with polyclonal anti-Myc Ab plus Cy3 anti-rabbit secondary Ab. (Bar = 1.5 μm.) (B) Dam1-GFP is associated with centromeric DNA in vivo, in an NDC10-dependent manner. CEN4 association was tested in wild-type or ndc10-1 cells at 37°C. Background signal was detected by using URA3-specific oligonucleotides. Total input, DNA extract of total lysate before immunoprecipitation; IP, DNA extract after immunoprecipitation.