Figure 3.

Ana3 is essential for basal body formation/maintenance in sensory neurons but not for centriole duplication. (A and B) WT (A) and Ana3 mutant (B) third antennal segments expressing GFP-PACT. In WT, this marker reveals the basal bodies at the base of each sensory bristle (arrow; magnified in insets). Basal bodies are undetectable in the mutant (mut). (C and D) WT (C) and Ana3 mutant (D) third antennal segments expressing the membrane marker mCD8-GFP. In WT, this marker reveals a cilium, a thin line extending into the bristle (C, arrow). Cilia are undetectable in the mutant. (E and F) WT (E) and Ana3 mutant (F) mitotic neuroblasts stained for DNA (blue), the centriole marker Asl (green), and the PCM protein Cnn (red). WT cells contain two centrosomes, whereas mutant cells frequently contain too few or too many centrosomes. The depicted mutant cell has five centrosomes with variable amounts of Cnn. The dimmest centriole (arrow) has the least Cnn. (G) Centrosome numbers (identified by colocalization of D-PLP and Cnn) in WT and Ana3 mutant prophase neuroblasts. Centrosomes were counted in a total of 32 WT and 39 mutant cells from four brains per condition. (H) Quantification of Cnn levels at metaphase centrosomes in WT and Ana3 mutant brain cells. Total fluorescence intensity was measured for a total of 40 centrosomes from four brains per condition. Bars: (A and B) 20 µm; (C–F) 10 µm.