|
Fig. S7 Brpf1-GFP displays PWWP domain-dependent chromatin association in zebrafish embryos. Zebrafish embryos were injected at the 1-cell stage with mouse full-length GFP-Brpf1 or GFP-Brpf1ΔPWWP plasmids, as also used for Fig. S1E, Figs 5 and 6, and fixed at mid-gastrula stages (7-9 hpf). GFP was visualized by immunofluorescence, as described for transfected HEK 293 cells (Figs 5 and 6), DNA was counterstained with DAPI, and embryos were analyzed by confocal microscopy (Zeiss LSM510 META). (A-C) Full-length Brpf1; (D) Brpf1ΔPWWP. (A) Interphase, (B) early mitosis, (C,D) late mitosis, with the two sets of sister chromosomes clearly separated. Full-length Brpf1 is localized at discrete sites of the chromatin of zebrafish cells both in interphase (A) and mitosis (B,C); compare with Fig. 5A and Fig. 6B for HEK 293 cells. (C,D) In contrast to full-length Brpf1 (C), Brpf1ΔPWWP lacking the PWWP domain is largely excluded from the chromatin (D); compare with Fig. 6D.