Figure 5.

Reb1 recruitment partially restores roadblock activity at centromeres lacking Cbf1. (A) Schematic of the mutations introduced on endogenous CEN8 that is flanked by a LacO array. The mutations on CDEI and the replacement of CDEI by the Reb1 binding motif are identical to those performed on CEN3 and presented in Figure 4. (B) RT-qPCR analysis of cenRNA5 and cenRNA8 expression levels. (C) Quantification of chromosome VIII mis-segregation in anaphase (percent of binucleate cells with a fluorescently labeled CEN8 signal in only one of the two nuclei). Error bars represent SD of three independent experiments; n = 200 cells for each experiment. P-values for (B) and (C) were determined using a two-tailed unpaired t-test (*P < 0.05; **P < 0.01; ***P < 0.001).