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Fig. 3. Histone macroH2A1.2 is not concentrated in X and Y chromatin of early pachytene XY oocytes. (Blue, DAPI; red, anti-SCP3 in A-D and anti-XLR in E-P; green, anti-macroH2A1.2 antibody; yellow/orange, co-localisation of anti-macroH2A1.2 with anti-SCP3 or anti-XLR; white, CREST antibody, which marks the centromeres). (A,B) In spread oocytes from XX mice, macroH2A1.2 is concentrated at the centromeric heterochromatin of all bivalents as well as at the non-centromeric end of one bivalent (arrow); this bivalent must be the X which has a PAR focus (Turner et al., 2001). (C,D) In this spread oocyte from an XY^Tdym1 mouse in which the X and Y are synapsed, the PAR focus of macroH2A1.2 is clearly visible (arrow). The thin asynapsed non-PAR axes of the X and Y are also distinguishable, but only the centromeric region of the X has a high concentration of histone macroH2A1.2. (E-H) In XX early pachytene oocyte squashes, anti-XLR antibody produces a granular but fairly uniform staining thoughout the nucleus. MacroH2A1.2, however, is concentrated in the regions where the centromeres are clustered. (I-L) In early pachytene XY oocytes, anti-XLR antibody now concentrates on the chromatin of the asynapsed X (arrow). (M-P) In late pachytene XY oocytes, macroH2A1.2 preferentially stains the X centromere (arrow in O) and the PAR focus. Arrow in N indicates the chromatin of asynapsed X. Scale bar: (A-D) 21 µm; (E-P) 7 µm.

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