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Fig. 7. Relationship between H3/K9 and DNA cytosine methylation in the male
pronucleus following transplantation into GV-stage or MII-stage oocytes.
Reconstructed oocytes were produced by transferring male pronuclei into
enucleated GV-stage oocytes, followed by a 3-hour culture to retrieve meiosis
(NT-GVB) or to inhibit meiotic resumption in the presence of IBMX (NT-GV).
Alternatively, the male pronuclei were transferred into enucleated MII-stage
oocytes (NT-MII), followed by culture for 3 hours in the presence of
nocodazole to prevent spontaneous activation. The antibodies directed against
H3/K9 methylation (MeH3K9) and DNA cytosine methylation (5-MeC) were detected
using the cy3-conjugated secondary antibody (red) and the FITC-conjugated
secondary antibody (green), respectively. The reconstruction experiments for
the NT-GVB, NT-GV and NT-MII oocytes were conducted three times. In total, 18
NT-GVB, 20 NT-GV and 18 NT-MII oocytes were reconstructed successfully and
examined for histone methylation. Similar patterns were observed for each type
of reconstructed oocytes; representative examples are shown.
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