Fig. 9. The incorporation of Nup93 into the NE occurs independently of the lamina. (A) Sperm pronuclei were assembled in egg extract for 90 min in either the presence (Delta 2+) or absence (Control) of Xlamin B₁ Δ2+. The nuclei were then collected on glass coverslips, extracted and fixed as described for Figure 5. The nuclei were stained with mAb 414 (F/GXFG, panels b, e, h) and anti-Nup93 (Nup93, panels c, f, i,). The distribution of mAb 414 was revealed with FITC-conjugated goat anti-mouse Ig while the distribution of anti-Nup93 was revealed with TRITC-conjugated goat anti-rabbit Ig. The distribution of chromatin in each image is revealed with DAPI (panels a, d, g). (B) Alternatively, sperm pronuclei were assembled in egg extracts over 90 min before Xlamin B₁ Δ2+ (Delta 2+) or control buffer (Control) was added to the incubation. Ninety minutes after the addition of the mutant protein, nuclei were processed for immunofluorescence as described in (A). The nuclei were stained with mAb 414 (F/GXFG, panels b, e, h), anti-Nup93 (Nup93, panels c, f, i) and DAPI (panels a, d, g). Scale bar = 20 µm.