Figure 3.

Association of minor satellite RNA with proteins of the chromosomal passenger complex. (A) In vitro transcribed minor satellite RNA (minsatF) was used in RNA pull-down experiments, followed by western blot (WB) analysis with the indicated antibodies. As controls, similar experiments were performed in presence of RNaseA, in absence of minor satellite RNA, or using an RNA not complementary to the biotinylated oligonucleotide used (minsatR). (B) RNA NChIP from nuclear extracts of asynchronously growing (AS) and nocodazole-arrested (NOC) MEL cells using antibodies against Aurora B, Survivin and H3K9Me3-S10ph. As a negative control, a similar experiment was performed in the absence of immunoprecipitating antibody (beads). Following DNaseI treatment, immunoprecipitated (IP) RNA and RNA from supernatant fractions (SUP) were analysed by RT-PCR, in reactions containing (+) or not (−) RT, for the presence of minor satellite RNA (minsat) and rDNA transcripts (rDNA).