Fig. 3. Treatment of injected stage VI oocytes with 50 nM LMB results in nuclear accumulation of the N-terminal region of Xp54 fused to GFP. (A) Oocyte nuclei, isolated 16 and 24 hours after injection with plasmids expressing NT-GFP, GFP or T7-RbAp48 (p48), viewed by phase microscopy (PC), direct fluorescence (FL) and confocal fluorescence (CF) are shown. (B) (left) A nucleus and cytoplasm from an oocyte injected with NT-GFP, treated with LMB and viewed for green and red fluorescence. (right) An immunoblot, using anti-myc IgG, of expressed NT-GFP present in the nucleus and cytoplasm 16 and 24 hours after plasmid injection. Injected oocytes were incubated with or without LMB. Protein equivalents of five nuclei (N) and one cytoplasm (C) were separated by SDS-PAGE. (C) (left) Translocation into the nucleus of anti-myc IgG from 20 ng injected into the cytoplasm of oocytes expressing GFP and NT-GFP with or without LMB. The anti-myc IgG was detected on slot blots with an HRP-conjugated anti-mouse IgG. (right) Translocation of anti-myc IgG injected into the cytoplasm of stage IV oocytes onto the RNP matrix of lampbrush chromosomes is shown. Anti-myc IgG was detected, after incubation of chromosome spreads from oocytes expressing NT-GFP or GFP, with FITC-conjugated anti-mouse IgG. Location of the chromosomal axis is seen by post-staining with DAPI.
