Nkd1 truncation alters subcellular localization and Dvl colocalization.

<p>(A, B) HEK-293 cells expressing HA/Flag-tagged Nkd1 (A) or Nkd1^C8 (B) stained with α-HA (green). Nkd1 accumulates in puncta (arrows), while Nkd1^C8 is diffusely distributed in the cytoplasm. Nkd1^C6 distributed in a pattern similar to Nkd1^C8 (not shown). (C, D) Cells co-expressing Dvl3 and wild-type (C) or C8 (D) Nkd1^GFP constructs. Nkd1-GFP (green, C) shows near complete colocalization (arrows) with Dvl3 (red, C′), while Nkd1^C8-GFP (D) accumulates in cytoplasmic aggregates surrounded by Dvl3 (arrows). Merged images are in C″, D″, DNA is blue in A–D. Similar results were observed in cells coexpressing Nkd1^C8-GFP and Dvl1 or Dvl2, and in cells expressing Nkd1^C6-GFP and Dvl1, Dvl2, or Dvl3 (not shown). (E) Salivary gland from wild-type third instar <i>Drosophila</i> larva stained with α-Dsh (red) showing diffuse and punctate staining. (F–F″) <i>A8-Gal4/UAS-Nkd^GFP</i> salivary gland stained with α-Dsh and imaged for GFP (F) and Dsh (F′; merged in F″). Fly Nkd^GFP relocalizes Dsh to perinuclear aggregates (arrows) similar to those observed with Nkd1^GFP/Dvl3 in C.