JCS021055 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. UAP56 is concentrated in RNA-splicing speckled domains and in the regions surrounding them. (A) HeLa cells were permeabilized, fixed and stained for both UAP56 and for SC-35. Confocal microscopy showed that UAP56 was present in RNA-splicing speckled domains, where it colocalized with SC35, a well characterized RNA-splicing speckled-domain protein, but was also present in the nucleoplasm. A low-magnification view of HeLa cells stained for UAP56 and SC-35. (B) A high-magnification view of one nucleus from a cell in A showing that UAP56, although present in speckled domains with SC-35, also is in nucleoplasmic regions surrounding speckled domains. Scale bars: 10 µm.

• Supplemental Figure S2 -

  Fig. S2. The mitotic choreography of UAP56. CaSki cells, grown on coverslips, were fixed and permeabilized. The cells were then immunostained for confocal microscopy using antibodies against UAP56 (green) and α-tubulin (red). DRAQ5 was used to localize DNA. Shown for each stage of mitosis are individual fluorescence channels, an overlay of a single section, a maximum intensity projection and a differential interference contrast (DIC) image. During metaphase, UAP56 was most concentrated at spindle poles, where some UAP56 remained until telophase. Through anaphase to telophase, the number of cytoplasmic foci increased as UAP56 appeared to leave the spindle poles. As the nucleus reformed UAP56 was redistributing to larger structures in the cytoplasm and ultimately to the nucleus. UAP56 was observed in midbodies through cytokinesis. Scale bar: 10 µm.