Fig. 5. H3/K9 methylation in male pronuclei that were transplanted into enucleated GV- or MII-stage oocytes. The reconstructed oocytes were produced by transferring the male pronucleus 6 hours after insemination into enucleated oocytes at (A) the GV stage (NT-GV) or (B) the MII stage (NT-MII), and were used for immunocytochemistry following culture for 3 hours with (for NT-GV) or without (for NT-MII) 0.2 mM IBMX. The antibody that recognizes the methylated lysine 9 on histone H3 (MeH3K9) was localized with FITC-conjugated secondary antibodies (green), and DNA was stained with DAPI (blue). Intact oocytes at the GV stage (GV) and MII stage (MII) that had not been enucleated are shown as controls. The reconstruction experiments for the NT-GV and NT-MII oocytes were conducted four and three times, respectively. In total, 23 NT-GV and 18 NT-MII oocytes were reconstructed successfully and examined for histone methylation. Similar patterns were observed for each type of reconstructed oocyte; representative examples are shown.