The N-terminus of EDC4 is not required for interaction with DCP1a.

Co-expression of NLS-EDC4 (panel a) and NLS-DCP1a (panel b) resulted in co-localization of the proteins to nuclear dots (panel c) in all cells that had both proteins in the nucleus. White arrows in a-c indicate a representative cell containing both NLS-EDC4 and NLS-DCP1a in nuclear dots. GFP-NLS-EDC4(630–1437) (panel d) was also able to recruit NLS-DCP1a (panels e, f) to nuclear dots in all cells that expressed both proteins. The C-terminus of EDC4 is sufficient to mediate interaction with CCHCR1. Co-expression of NLS-EDC4 (panel g) and CCHCR1-Myc (panel h) resulted in co-localization of the proteins in nuclear dots (panel i) in all cells that expressed both proteins. The C-terminus of EDC4 (GFP-NLS-EDC4(935–1437)) (panel j) was sufficient to recruit CCHCR1-Myc (panels k, l) to nuclear dots. CCHCR1-Myc co-localized with GFP-NLS-EDC4(935–1437) in 96+/-4% of cells expressing both proteins. White arrows indicate the location of representative cells that expressed NLS-EDC4 and CCHCR1-Myc (g-i) and NLS-EDC4(935–1437) and CCHCR1-Myc (j-l). Human serum was used to detect NLS-EDC4 in a and g. Rabbit antiserum was used to detect NLS-DCP1a in b and e. Rabbit anti-Myc antiserum was used to detect CCHCR1-Myc in h and k. Mouse monoclonal anti-GFP antibody detected GFP-NLS-EDC4(630–1437) and GFP-NLS-EDC4(935–1437) in d and j respectively. Merge of panels a and b, d and e, g and h, and j and k is shown in c, f, i and l respectively. DAPI staining (blue) indicates the location of nuclei in c, f, i and l.