Stable inheritance of CENP-A at centromeres through cell divisions as well as transgenerationally. (A) Schematic depicting pulse-chase assay based on CENP-A-SNAP labeled with a fluorescent dye (TMR-Star). Quantification of CENP-A-SNAP intensity through time reveals a half-life equal to that of the cell cycle, indicating quantitative inheritance. (B) Schematic of cell cycle–coupled dilution of centromeric chromatin determined by CENP-A-SNAP pulse-chase assay. A.U, arbitrary units. (C) Experimental setup to determine dynamics of preincorporated CENP-A through meiotic arrest in mice. Endogenous CENP-A coding region is flanked by loxP sites. The Cre recombinase is expressed under Gdf9 promoter, which is active only in early stage oocytes at birth. Cre-mediated excision of CENP-A at this stage ensures that there is no further CENP-A assembly until oocyte maturation at 11–14 mo. CENP-A measurements in fully grown oocyte at birth and ovulated oocytes after 11–14 mo shows retention of CENP-A (red dots), which was assembled before birth.