Fig. S5.

Experimental control for double immunolabeling of centromeres using both rabbit anti-CENH3 and rabbit anti-KNL2 antibodies, on WT roots. The table indicates the followed protocol. The three (blue, red, and green) splitted channels (using ImageJ) are merged on the fourth panel (Right). (A) Colocalization of CENH3 and KNL2 in interphase. When KNL2 antibodies are incubated after (B) or before (C) CENH3 antibodies in metaphase, the absence of KNL2 labeling attests for the absence of artefactual fluorescence cross reactions. No fluorescence is observed in interphase nuclei, in the green channel, after either anti-CENH3 (D) or anti-KNL2 (E) preincubation, confirming the specificity of the antibodies. DAPI staining in blue. (Scale bars, 2 μm.)