Fig. 4. AKAP95 is more strongly anchored in NT pronuclei than in ICSI or parthenogenetic pronuclei. (A) Mouse one-cell-stage ICSI, parthenogenetic (Part) and NT embryos were double immunolabeled using anti-lamin-B (red) and anti-AKAP95 (green) antibodies. DNA was labeled with Hoechst 33342. Only nuclei are shown. FPN, female pronucleus; MPN, male pronucleus. Scale bars, 20 µm. (B) Mouse one-cell-stage ICSI, parthenogenetic (Part) and NT embryos, and mouse cumulus cell nuclei, were extracted with 0.1% Triton X-100, 1 mg ml^-1 DNase I, 100 µg ml^-1 RNase A together with 100 mM (upper two rows) or 300 mM (lower two rows) NaCl prior to fixation and double immunofluorescence analysis of lamin B (red) and AKAP95 (green). DNA is labeled blue with Hoechst 33342. Only nuclei are shown. Scale bars, 20 µm. (C) Relative proportions of unextracted lamin B, AKAP95 and DNA in nuclei of embryos or cumulus cells extracted as in (B). Fluorescence labeling intensity of each marker after in situ extraction as in (B). Reference value (100%) represents immunofluorescence labeling intensity in unextracted embryos. n10 embryos per marker per treatment.

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