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Fig. 4. SCP3 is located beneath associated sister kinetochores at the inner domain of metaphase I centromeres, showing a complex `double cornet'-like arrangement, and colocalizing with the cohesin Rad21. SCP3 is labelled in green, and centromeres (ACA), CENP-A, CENP-E, INCENP, Rap1 and Rad21 are labelled in red. In merged images colocalization regions appear in yellow. SCP3 appears as a T- or Y-like structure in centromeres viewed from the side (A,C,E,F,H,I), but as two closely associated rings when centromeres are viewed from the top (B,D,G,J). SCP3 appears beneath the ACA and CENP-A signals with which there is some degree of colocalization (A,C). The ACA, and the pair of CENP-A dots representing sister kinetochores, appear surrounded by the SCP3 rings (B,D). CENP-E appears as a single plate separated from the underlying SCP3 signal (E). By contrast, SCP3 colocalizes at the inner centromere domain with INCENP (F) and Rad21 (I). In this sense, Rad21 is the only protein appearing as two closely associated rings colocalizing with SCP3 when centromeres are viewed from the top (J). Sister proximal telomeric sequences, as detected with the anti-Rap1 antibody, are detected as a pair of dots partially colocalizing with the middle region of the T-shaped SCP3 signal (H). Scale bar: 1 µm.
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