Fig. 4. AKAP95 associates with maternal chromosomes at first mitosis, whereas mCAP-D2 is recruited to both chromosome complements. (A) Sperm were labeled with Hoechst 33342 (blue) before ICSI into oocytes. Resulting ICSI embryos were fixed at the PN stage, total DNA was labeled with PI (red) and AKAP95 localized by immunofluorescence (green). Merge image shows AKAP95 labeling restricted to the FPN (yellow). (B,C) Mitotic zygotes produced by ICSI of Hoechst-labeled sperm as in A were fixed and AKAP95 (B) and mCAP-D2 (C) were examined by immunofluorescence. (D) Mitotic embryos were also double-labeled with anti-AKAP95 mAb47 and anti-hCAP-D2 antibodies. `F' and `M', female and male chromatin, respectively; PB, polar body. Bars, 20 µm.