Figure 2

Immunostaining of centromeric proteins and multicolour fluorescence in situ hybridisation (FISH) using α-satellite DNA probes. (a) Centromere regions in hTERT-RPE1 cells were detected through CREST staining (red) and were combined with CRISPR imaging targeting α-satellite DNA (green). DNA was counterstained using Hoechst 33342 (blue). Two centromere regions enclosed by white boxes (1 and 2) are enlarged, and their line profiles, indicated by white dashed lines, are shown by graphs. (b) hTERT-RPE1 cell chromosomes were distinguished using 24 × Cyte probes, and individual chromosomes were detected using different colours (left). The hybridisation of DNA oligo probes with the targeting site of sgRNA 3 (α-satellite 3) showed green signals at the centromeres (right). Green arrows indicate chromosomes with signals of the α-satellite 3 probe, and enlarged chromosome images are also presented. (c) Percentage of cells showing α-satellite DNA signals on chromosomes 1–22 and X. Twenty cells were analysed.