Eic1 and Eic2 promote normal levels of association of Cnp1CENP-A assembly factors with centromeres. (a,b) Mis18-GFP and Mis16-GFP association with centromeres is entirely dependent on Eic1, and (e,f) partly dependent on Eic2. (c) Scm3-GFP association with centromeres is dependent on Eic1, and (g) largely independent of Eic2. (d) Mis6-HA association with centromeres is partly dependent on Eic1, and (h) Eic2. qChIP analyses of (a,e) Mis18-GFP, (b,f) Mis16-GFP, (c,g) Scm3-GFP and (d,h) Mis6-HA association with centromeres in the indicated strains when grown at permissive (25°C) versus restrictive temperature (36°C) for 8 h (a–d); or at 32°C (e–h). Enrichment of cc2 DNA relative to the act1 locus is presented in (a–d). Enrichment of cc2 or cc1/3 DNA relative to the act1 locus is presented in (e–h). Error bars represent standard deviation between at least three biological replicates.