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Fig. 4. Pin1-/- PGCs do not undergo cell cycle arrest and apoptosis. (A,C,E) Wild type; (B,D,F) Pin1-/-. (A-F) Confocal images of 12.5 dpc male (XY) gonads after wholemount immunohistochemistry. (A,B) Nearly all PGCs (green, round cells) are positive for the Ki67 antigen (red, arrowheads) in wild-type (A) and Pin1-mutant (B) gonads, indicating that PGCs are actively cycling in the presence and absence of Pin1. (C,D) Phosphohistone H3 (green, arrows) identifies PGCs (red, round cells) in mitosis in wild-type (C) and Pin1-/- (D) gonads. No accumulation of phosphohistone H3-positive cells is seen in Pin1-/- gonads, indicating that Pin1-/- PGCs are not arrested in mitosis. (E,F) Wild-type (E) and Pin1-mutant (F) PGCs (green, round cells) are not positive for the apoptosis marker LysoTracker (red), indicating that Pin1-/- PGCs do not undergo increased apoptosis.