FIGURE 1:

α-Satellite associations with nucleoli are reduced upon depletion of Ki-67 but not nucleolin. (A) Immuno-FISH analyses of the association between α-satellite DNA from chromosome 17 (αSat 17, red) and fibrillarin (green) was performed in HeLa cells transfected for 72 h with in vitro–diced esiRNAs targeting luciferase, Ki-67, or NCL (black bars) or with synthetic duplex siRNAs (checked bars) targeting Ki-67 or a scrambled sequence control (si-Scr). The percentage of αSat 17 alleles colocalized with fibrillarin is indicated, with means and SD error bars from three replicate experiments. p values comparing association frequencies in cells treated with esi-Luc (669 alleles) vs. esi-Ki-67 (573 alleles) and si-SCR (540 alleles) with si-Ki-67 (567 alleles) are indicated. (B) Representative FISH images of cells analyzed in A. Scale bar, 10 μm. (C) Immunoblot analyses of cells from A. Blots were probed with antibodies recognizing Ki-67, α-tubulin (loading control), NCL, or fibrillarin (loading control), as indicated.