Ana3 is required for centriole structure and cohesion. (A–E) WT (A and C) and Ana3 mutant (mut; B, D, and E) primary spermatocytes expressing GFP-PACT (green) stained for tubulin (red) and DNA (blue). (A and C) WT primary spermatocytes contain two v-shaped centriole pairs (A; magnified in inset), and the centrioles in each pair remain tightly associated until the end of meiosis I (C). (B) In the Ana3 mutant, centriole numbers are reduced, and the centrioles (magnified in the inset) are almost never paired. (D and E) Prematurely separated centrioles nucleate asters during meiosis (D), leading to multipolar spindles (E). (F) Centriole numbers in WT, Ana3 mutant, and Ana3 mutant expressing Ana3-GFP (rescue) G2 primary spermatocytes. Centrioles were counted in a total of 163 WT, 182 mutant, and 131 rescue cells from at least seven testes per condition. (G) Quantification of centriole intensity (marked by GFP-PACT) in WT and Ana3 mutant meiosis II spermatocytes. A total of 71 centrioles were measured from five testes per condition. (H) A WT centriole and six mutant centrioles (marked with GFP-PACT) from spermatocytes in prophase of meiosis II. Bars, 10 µm.