Fig. 4. Anillin and Pav-KLP fail to be recruited to ring canals in orbit⁶ mutants. (A-E) Germaria stained with anti-Anillin (red) and anti-HtsF (green) antibodies. (A) Wild-type cyst showing Anillin recruited to the newly formed contractile rings (arrow) before the initiation of fusome plug formation. (B) Wild-type cyst with Anillin present in the constricting and migrating ring canals. (C) Wild-type 16-cell cyst that has completed the four rounds of divisions. Anillin is present in the ring canals situated along the fusome. Only eight ring canals and the corresponding fusome part are visible; the others are out of focus. (D,E) orbit⁶ germaria that contain one and two germline cysts respectively, each with several fusome pieces. No Anillin-containing ring canal-like structures are visible. However, as with wild-type, the interphase nuclei of mutant cysts contain Anillin. Scale bar: 5 µm. (F-H) Germaria were stained with anti-Pav-KLP (green) and anti-HtsF (red) antibodies. (F) Wild-type stem cell and cystoblast interconnected through a Pav-KLP-containing ring canal at the time of fusome partition. The nuclei of these cells contain elevated levels of Pav-KLP, suggesting that these cells are in interphase. (G) Pav-KLP recruited to the ring canals surrounding the pre-existing fusome in wild-type germline cysts. (H) orbit⁶ germline cyst with an abnormal fusome and Pav-KLP accumulated in a single ring canal-like structure (broad arrow). Fusome pieces seen at the posterior of this cyst are of different sizes and show little or no Pav-KLP staining. Scale bars: 5 µm in F,G; 10 µm in H.

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