Fig. 5

ImmunoFISH detection of somatic nuclei prepared with the non-enzymatic in-solution procedure. A Top row: single channel (monochrome) images of a 3D-rendered barley somatic nucleus. The CENH3 protein is shown in white (immunolabelling) and the barley centromere-specific G+C-rich satellite is shown in green (FISH). The chromatin is counterstained with DAPI (blue). Centromeric signals are shown in surface mode. The enlarged bottom image shows all channels merged into a single image. Bars = 5 µm. B Top image: an enlargement of the centromeres shown in the bottom of A (bar = 5 µm); bottom image: enlargement showing the structure of a single barley centromere (bar = 1 µm). C Single channel (monochrome) and merged immunoFISH images of a 3D-rendered wheat somatic nucleus. Active centromeres are labelled with an anti-CENH3 antibody (white on merge). The centromeric retrotransposon of wheat (CRW, green on merge) and the telomeric repeat sequences (TRS, magenta on merge) are detected by FISH. The chromatin is counterstained with DAPI (blue on merge). Bar = 5 µm. D Single channel (monochrome) and merged immunoFISH images of a 3D-rendered somatic nucleus of a wheat × barley F1 hybrid. Active centromeres are labelled with an anti-CENH3 antibody (white on merge). The centromeric retrotransposon of wheat (CRW, orange on merge) and the barley centromere-specific G+C-rich satellite (green on merge) are detected by FISH. The chromatin is counterstained with DAPI (blue on merge). Bar = 5 µm