Ecdysone signaling modulates NC removal. (A) Schematic diagram of NC encapsulation by 4–5 AFCs during late oogenesis (stage 11 to late stage 12). (B′–C″) Immunostained egg chambers depicting follicle cells in the act of enveloping the NCs. Stage 11 egg chambers exhibit small fingerlike projections (B′ and B″). Stage 13 egg chamber where the NC is completely enveloped by the overlying follicle cells (C′ and C″). Nuclei are shown in red, and follicle cells are marked with mCD8GFP in green. (D¹–E²) Stagewise activation of ecdysone signaling in the AFCs during late oogenesis. EcRE-lacz is marked in green, and nuclei are marked in red. Stage 11 egg chamber (D¹ and D²) and stage 12 egg chamber (E¹ and E²). (D²and E²) are LUT images of EcRE-lacz. (F′–I″) Stage 14 egg chambers of indicated genotypes. Persisting NC nuclei are marked by a yellow arrowhead. DAPI is in red, and Armadillo is in green. Images show control (F′ and F″), EcR^RNAi (G′ and G″), EcRB1^DN (H′ and H″), and Usp^RNAi of genotype GR1-Gal4/Usp^RNAi (I′ and I″). (J) Quantification of persisting NC nuclei in stage 14 egg chambers of indicated genotypes. “n” stands for the number of egg chambers analyzed. (K) Alternative quantification of PN in stage 14 egg chambers. The number of PN was categorized into bins of 0 PN, 1–3 PN, 4–6 PN, 7–10 PN, 11–15 PN. The percentage of stage 14 egg chambers exhibiting PN in each bin was calculated. n represents the number of egg chambers analyzed. Error bars represent the SEM, and **** represents the level of significance P < 0.0001.