Figure 4. Depletion of I3 induces re-localization of SDS22 to kinetochores.

1. Neither SDS22 nor I3 colocalize with PP1γ at metaphase kinetochores under control conditions. Stable HeLa cell lines expressing GFP fusions of indicated proteins were fixed and DAPI-stained. SDS22-GFP is expressed under its own promoter in BAC transgenic cells. For localization images of GFP-I3-expressing cells in all mitotic stages see also Supplementary Fig S2. Representative confocal images. Scale bar, 10 μm.
2. Expression levels of GFP fusions relative to endogenous proteins in Western blots stained with specific antibodies. Actin served as a loading control. Asterisks denote non-specific bands.
3. SDS22 re-localization upon I3 depletion. SDS22-GFP cells were treated with indicated siRNAs and processed as in (A). Representative images. Note the SDS22-GFP signal at kinetochores in I3, but not control-depleted cells. Scale bar, 10 μm.
4. Depletion efficacy for individual siRNAs as determined by Western blot.
5. Quantification of (C). Error bars indicate s.d. of three independent experiments with 25 cells per condition. ***P < 0.001.

Source data are available online for this figure.