Fig. 6. Involvement of protein synthesis and gene expression in H3/K9 methylation in mouse embryos. (A) Cycloheximide (50 µg/ml) was added at the time of insemination, and H3/K9 methylation (MeH3K9) was examined 12 hours post-insemination. (B) -Amanitin (25 µg/ml) was added 4 hours after activation of the androgenetic embryos, and H3/K9 methylation was examined 32 hours post-activation. The polyclonal antibody that recognizes methylated H3/K9 was localized with FITC-conjugated secondary antibodies (green), and the DNA was stained with DAPI (blue). The experiments using cycloheximide and -amanitin were conducted three and two times, respectively, and similar results were obtained. 21, 12, 18 and 11 embryos were examined in the cycloheximide (+), cycloheximide (-), -amanitin (+) and -amanitin (-) groups, respectively, and representative examples are shown.