Fig. 2.

The integrin heterodimer pair αPS3/βPS is required in follicle cells for engulfment. (A-L) Egg chambers from starved flies are labeled with DAPI (cyan), cleaved anti-Dcp-1 (yellow) and anti-Dlg (red) to visualize membranes. Antibody against cleaved Dcp-1 specifically marks the dying germline and engulfed material. (A-C) Control (GR1-GAL4, G89/TM6B) egg chambers show normal death and engulfment. (A) Healthy egg chambers do not label with Dcp-1. (B,B′) A phase 3 dying egg chamber shows several Dcp-1-positive particles (arrows) inside the follicle cells (FCs), indicating engulfment. (C) A phase 5 dying egg chamber shows little remaining germline, and few Dcp-1-positive particles still inside the FCs. (D-F) Egg chambers expressing αPS3^dsRNA specifically in the FCs are normal when healthy (D) but have little to no particle uptake in phase 3 (E,E′; arrows). (F) Loss of αPS3 in the FCs results in lingering germline material by phase 5. (G-I) Egg chambers expressing βPS^dsRNA specifically in the FCs are similar to those with loss of αPS3, with few particles (H,H′; arrows) and lingering germline material (I). (J-L) draper^−/− egg chambers (Etchegaray et al., 2012) are normal when healthy (J) but have little to no particle uptake in phase 3 (K,K′; arrows). (L) Loss of draper in the FCs results in lingering germline material. (M) Quantification of unengulfed germline. (N) Quantification of the number of Dcp-1-positive particles in phases 1-4. All data are mean±s.e.m. At least three egg chambers were quantified for each genotype, phase and quantification method in M,N. For M, 238 egg chambers were quantified for the control (a mixture of GR1-GAL4, G89 and sibling controls); 57 for αPS3^dsRNA; 110 for βPS^dsRNA; and 121 for draper^−/−. For N, 103 egg chambers were quantified for the control, 33 for αPS3^dsRNA, 33 for βPS^dsRNA and 44 for draper^−/−. One-way ANOVA and Bonferroni-Holm post-hoc tests were performed: ***P<0.005, *P<0.05, n.s., non-significant. Scale bar: 50 µm.