(A) Representative images of Ska3-Ndc80 Proximity Ligation Assay (red), with additional immunostaining of Borealin (cyan) and tubulin (green). Top – prometaphase cells, middle – metaphase cells, bottom – control metaphase cells.
(B) Centromeres in prometaphase or metaphase cells, as identified by Borealin immunostaining, were scored based on the proximity to Ska3-Ndc80 PLA signal (mean ± SD; N > 100 centromeres from four cells). Cells with only Ska3 probe were used as a control. N/S – p > 0.05, * - p < 0.05, *** - p < 0.0001. (Student’s t-test)
(C) Centromeres in prometaphase or metaphase cells, as identified by Borealin immunostaining, scored based on the number of proximal Ska3-Ndc80 PLA signals (mean ± SD; N > 100 centromeres from four cells). N/S - p > 0.05, * - p < 0.05, *** – p < 0.0001. (Student’s t-test)