Extended Data Fig. 4: Overexpressing NCAPD3 or NCAPH2 did not rescue centromere stretching.
From: Condensin dysfunction is a reproductive isolating barrier in mice
Hybrid oocytes expressing eGFP-NCAPD3 (a) or NCAPH2-eGFP (b) derived from domesticus or spretus were fixed at prometaphase I and stained for TOP2A (a major satellite marker, see Extended Data Fig. 8b) and eGFP. The length of major satellites was quantified; each dot in the graph represents a single centromere (a, n = 752, 683 and 743 centromeres for control, + spretus eGFP-NCAPD3 and + domesticus eGFP-NCAPD3, respectively; b, n = 464, 294 and 451 centromeres for control, + spretus NCAPH2-eGFP and + domesticus NCAPH2-eGFP, respectively). NCAPH2-eGFP was not able to localize on the chromosome probably because of the competition with endogenous NCAPH229. The enhanced centromere stretching upon overexpressing NCAPH2-eGFP implies that NCAPH2-eGFP sequesters other condensin II subunits in the cytoplasm, reducing the functional condensin II pool. Images are maximum intensity z projections to show all chromosomes or optical slices to show individual chromosomes; red line, mean; scale bars: 5 µm.
