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. 2018 Nov 26;7:e40325. doi: 10.7554/eLife.40325

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.

Figure 11.

Figure 11—figure supplement 1. Localization of HEC1 in HeLa cells depleted of SF3A2 and PRP31 and stained for HURP.

Figure 11—figure supplement 1.

(A) Representative control (mock), SF3A2 RNAi and PRP31 RNAi cells fixed after pre-extraction with Triton X-100 (see Materials and methods), and stained with DAPI (DNA, blue), anti-HURP (red) and anti-HEC1 (green) antibodies.