Figure 2

Accumulation of Dsh in the nucleus in the absence of nuclear export. (a-d) Dsh-GFP RNA (0.7 ng) was injected into two animal blastomeres of 4–8 cell embryos. Animal caps were excised at stage 10 and then left (a) untreated or (b) treated with 10 mM NEM or (c,d) 50 ng/ml leptomycin B (LMB), fixed and examined for GFP fluorescence. (a) Dsh-GFP is mainly localized to vesicular structures in the cytoplasm. In the presence of (b) NEM or (c) LMB, Dsh-GFP accumulates in the nucleus, as supported by (d) DAPI staining of nuclei in the same field as in (c). Nuclear staining is marked by arrowheads (c,d). (e,f) The Ds3 construct, lacking amino acids 334–381, remained in the cytoplasm in the (e) absence or (f) presence of NEM.