Fig. 2. Subito is required during anaphase for the localization of Polo and Aurora B but not Fascetto or Pavarotti. All images are stained for DNA (blue) and, except for A and B, tubulin (green). Polo (green) colocalizes with Subito (red) at the midzone in wild-type anaphase (A) but not in sub¹/sub¹³¹ mutants (B). (C) Aurora B (red) is localized on the chromosomes during metaphase (not shown) and during anaphase it moves off of the chromosomes and localizes on the spindle midzone. (D) In sub¹/sub¹³¹ mutant anaphase, Aurora B protein is still present on the anaphase chromosomes but not at the midzone. (E,F) Although usually more concentrated near the chromosomes in sub mutant anaphases, Aurora B and Incenp (red) often appeared to have spread out along the microtubules; they were never concentrated in the midzone. In panel E, there was no tubulin staining. Bars, 5 µm.