Fig. 5

Behavior of TF stacks during ovarian formation. Confocal images (A–F) and zoomed-in for outlined areas (A’–F’) of the ovaries of indicated developmental stages. Vasa (green), α-Spec/phalloidin (red), DNA (white), and PH3 (blue) staining. A, A’ PGC mass elongates along the A/P axis during mid-L4. B, B’ Terminal filament formation and branched fusomes breakdown occur during early wL4. C, C’ PGC mass breakdown into 2–6 clusters and pre-ovariole formation during late wL4. D, D’ Pre-ovarioles grow and migrate. E, E’ Pre-ovarioles continue to grow and migrate with ovariole stalks pointing toward the oviduct. F, F’ Pre-ovarioles develop into two segments (indicated by a yellow rectangle). Confocal images of pre-ovarioles/ovarioles of wL4 (G), 0 h pupa (H), 6 h pupa (I), 12 h pupa (J), 18 h pupa (K), 24 h pupa (L), 30 h pupa (M), 0 h adult with (N) or without (O) muscle sheath, 24 h adult (P), and 72 h adult (Q) with α-Spec/phalloidin (red) and DNA (white) staining show the development of terminal filament (TF), ovariole stalk (OS), germarium (G), and the primary follicle (F). White arrows in B–F mark the oviducts. Yellow dashed lines in B’–D’ mark one TF. Yellow arrowheads in G–Q mark the TF position. Yellow dash line in H marks the developing OS. Yellow lines in I–Q mark the OS. Scale bars in A (for A–F), A’ (for A’–F’), and Q (for G–Q) indicate 50 µm