JCS02736 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. An antibody against OsCenH3 can precisely recognize rice centromeres in mitosis and meiosis. (A) The amino acid sequence of centromere-specific histone H3 variant of O. sativa (OsCenH3), Zea mays (ZmCenH3, CF647791) and A. thaliana (AtCenH3, AY086361). Conserved amino acids are highlighted. The peptide sequence used for the antigen is underlined. (B) Western blot analysis using anti-OsCenH3. Crude extracts from adult leaves and nuclear extract from seedlings were loaded into lanes 1 and 2, respectively. (C and D) Indirect immunofluorescence of OsCenH3 for mitotic chromosomes at prometaphase (C) and for meiotic chromosome at metaphase I (D). Bar, 10 mm. (E-G) FISH analysis of Cent-O centromeric repeats (E) by immunofluorescence of OsCenH3 (F) on a mitotic chromosome spread. Both signals completely overlap (G). In G, chromosomes are counter-stained with DAPI (blue).

• Supplemental Figure 2 -

  Fig. S2. Specific localization of PAIR2 on centromeric regions is already observed at pachytene. (A) The left panel indicates spread chromosomes in the pachytene meiocyte with destructed nucleus. The right panel indicates a merged image of PAIR2 (green) and OsCenH3 (red). (B) A magnified view of the box in (A). An island of PAIR2 signal was clearly detected at the centromeric and peri-centromeric regions. Bar, 5 mm.