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Figure 4


Fig. 4. Histone H4 hyper-acetylation and de novo H2A ubiquitylation characterise the stages before histone removal. (A-C) Double immunostaining of core histones and acetylated histone H4 on squashed testes of protamine-eGFP flies. (D-F) Double immunostaining of core histones and mono-ubiquitylated histone H2A on squashed testes of protamine-eGFP flies. (A,D) Histones are abundant in round spermatids (arrows), whereas the staining decreases in early canoe stage spermatids (arrowhead in A). In later stages when protamines are present (double arrow in C), histones are no longer detectable. (B) Nuclei of round spermatids show a faint staining of acetylated histone H4 (arrow). In early canoe stage spermatids, acetylation of histone H4 increases (arrowhead) and has vanished completely from the late canoe stage onwards. (C,F) Protamine-eGFP expression starts in late canoe stage spermatids (double arrow in C). (E) Mono-ubiquitylated histone H2A is detectable in spermatids with round nuclei (arrow). Mono-ubiquitylated histone H2A is no longer detectable during the canoe stage and in later stages when protamines are present (double arrow in C). Bars, 5 µm.





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