Immunofluorescence analysis of the role of RNA in the assembly of centromere proteins on HeLa cell metaphase chromosomes. (A,B) Centromere binding of CENPA (Aii,Bii; red) and CENPE (Aiii,Biii; green) was not affected by RNase (which targets both single- and double-stranded RNA) treatment of the cells. (C–J) Centromere localization of human CREST antiserum (CR, which recognizes CENPA and CENPB) was not affected by RNase treatment of the cells (Cii–Jii; red). A similar treatment resulted in the complete delocalization of HP1-alpha (Ciii,Diii; green), INCENP (Giii,Hiii; green), and survivin (SURV) (Iiii,Jiii; green). This treatment caused a significant but not complete delocalization of CENPC1 from the centromere (Eiii,Fiii; green). Plus and minus signs indicate with and without RNase treatment, respectively. (i) Merged images of ii and iii.
