Fig. 1. Distribution of AKAP95 in mouse somatic cells, gametes and embryos. (A) Immunofluorescence analysis of AKAP95 in mouse cumulus cells and in situ-prepared nuclear matrices after methanol fixation, using an affinity-purified anti-rat AKAP95 antibody. DNA was counterstained with Hoechst 33342. (B) HeLa cells and mouse cumulus cells were immunoblotted using anti-AKAP95 antibodies with or without a competitor AKAP95(387-692) peptide. (C) Immunoblotting analysis of AKAP95 in MII oocytes, sperm, DTT-treated decondensed sperm `halos' and PN embryos. Protamines were immunoblotted to demonstrate availability of chromatin sperm antigens on blots.

(D) Immunofluorescence distribution of AKAP95 in MII oocytes and in vitro cultured preimplantation embryos. Arrowheads point to MII chromosomes. Arrows point to a metaphase blastomere. (E) MII oocytes were activated with either 10 mM SrCl₂, 10 µg/ml cycloheximide (CHX) or 10 mM SrCl₂ together with 5 µg/ml actinomycin D. Activated oocytes were fixed at the PN stage and labeled using anti-AKAP95 antibodies. Insets, MII and SrCl₂-activated oocytes were immunoblotted using anti-AKAP95 antibodies. DNA was stained with Hoechst 33342. FPN and MPN, female and male pronucleus, respectively. PB, polar body. Bars, 10 µm (A); 20 µm (D,E).