Fig. 5: The BUB1 kinase-H2A-pT121 pathway recruits pericentromeric factors to telomeric PMsat.

a Schematic of two pathways recruiting pericentromeric factors. b P. maniculatus meiosis I oocytes treated with 5-Itu were fixed at meiosis II and stained for HEC1 and H3-pT3. n = 16, 5, and 10 cells from three independent experiments for control, 0.5 µM 5-Itu, and 1µM 5-Itu, respectively. c P. maniculatus meiosis I oocytes expressing dCas9-EGFP with gRNA targeting PMsat were treated with 5-Itu, fixed at metaphase I, and stained for HEC1 and Survivin. Survivin levels at telomeric PMsat were quantified; each dot represents one chromosome; n = 26 and 23 cells from three independent experiments for control and the 5-Itu-treated group, respectively; red line, median. d P. maniculatus meiosis I oocytes treated with BAY-1816032 were fixed at metaphase I and stained for H2A-pT121 and HEC1. H2A-pT121 levels on chromosomes were quantified; each dot represents one oocyte; n = 13 and 11 oocytes from three independent experiments for control and the BAY-1816032-treated group, respectively; unpaired two-tailed Mann-Whitney test was used for statistical analysis; red line, median. e P. maniculatus meiosis I oocytes expressing dCas9-mCherry with gRNA targeting PMsat were treated with BAY-1816032, fixed at metaphase I, and stained for Survivin and HEC1. Survivin levels at telomeric PMsat were quantified; each dot represents an individual chromosome; n = 16 and 10 oocytes from three independent experiments for control and the BAY-1816032-treated group, respectively; unpaired two-tailed Mann-Whitney test was used for statistical analysis; red line, median. f P. maniculatus meiosis I oocytes expressing dCas9-mCherry with gRNA targeting PMsat were treated with BAY-1816032, fixed at metaphase I, and stained for MCAK and HEC1. MCAK levels at PMsat were quantified; each dot represents an individual chromosome; n = 122 and 141 chromosomes from four independent experiments for control and the BAY-1816032-treated group, respectively; unpaired two-tailed Mann-Whitney test was used for statistical analysis; red line, median. Orange asterisks, internal PMsat; yellow asterisks, telomeric PMsat; exact P values are in the graphs except for ****P < 0.0001; scale bars, 5 µm. Source data are provided as a Source Data file.