Figure 11. SF3A2 and PRP31 are required to prevent diffusion of HEC1 from kinetochores of HeLa cells.
(A, B) Prometaphases and prometaphase-like figures were fixed after pre-extraction with Triton X-100 (see Materials and methods) and stained with DAPI (DNA, blue) and either CREST (red) and anti-HEC1 (green) antibodies (A) or anti-tubulin (red) and anti-HEC1 (green) antibodies (B). Boxes delimit regions shown as enlargements in the rightmost panels. In SF3A2 and PRP31 RNAi cells the anti-HEC1 antibodies stain filaments protruding from the CREST signals (A); these filaments are also stained by anti-tubulin antibodies (B). The middle panels in A show two cells, C1 and C2. In C1, HEC1 is mostly associated with the kinetochores, showing only short protrusions from the CREST signals; in C2, the bulk of HEC1 is no longer on kinetochores but is instead associated with filamentous structures. Scale bar, 5 μm.