Extended Data Fig. 5: NCAPG2 amino acid sequences are mostly conserved between domesticus and spretus.

a, The entire amino acid sequences of NCAPG2 from H. sapiens, M. musculus domesticus (musculus), and M. spretus were aligned. Residues diverged between domesticus and spretus were highlighted in yellow. Although there are several residues diverged between domesticus and spretus NCAPG2, these differences are probably not part of the hybrid incompatibility, because both of them showed similar localization pattern and efficiently rescued centromere stretching when overexpressed in hybrid oocytes (Fig. 2c and Extended Data Fig. 7b). NCAPG2 antibodies used in this study (Bioss bs-7721R and Bethyl A300-605A) were raised against human NCAPG2 fragments. Antigen sequences used for the antibody productions were highlighted in light green for the Bioss antibody and light blue for the Bethyl antibody. b,c, Part of the amino acid sequences of human NCAPD3 (b) and NCAPG2 (c) that were used as the antigens (highlighted in grey) to produce the Bethyl A300-604A and A300-605A antibodies were aligned with NCAPD3 and NCAPG2 sequences from M. musculus domesticus, M. spretus and M. spicilegus. A single residue diverged among mouse species were highlighted in yellow. Since the sequences are basically conserved across the mouse species for the region corresponding to the antigen sequences (a-c), it is unlikely that the distinct NCAPG2 and NCAPD3 staining patterns among these mouse species are due to the differential efficiency in antibody recognition (Figs. 2b, 3a, 5b, d and Extended Data Fig. 2a, 6b, d).