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Meiosis was blocked at leptotene stage in both male and female germ cells of Wt1R394W/R394W gonads. Immunofluorescent staining of SYCP3 (red) and γH2AX (green) were performed on chromosome spreads of control (Wt1+/R394W; A–F: female; J–L: male) and Wt1R394W/R394W (G–I: female; M–O: male) gonads which were dissected at E12.5 and cultured in vitro for 3 days. The gender of the embryos was confirmed with PCR using Sry primers.
