Ecdysone pathway facilitates NC removal through Eip93F (E93). (A ¹ –D ³ ) Expression of Eip93F during different stages of late oogenesis: stage 10 (A¹–A³), stage 11 (B¹–B³), stage 12 (C¹–C³), stage 13 (D¹–D³). The expression of Eip93F starts after stage 11 only in the AFCs. White and magenta arrowheads mark AFC nuclei that express Eip93F. Eip93F is in green, and DAPI is in red. (E¹–H³) Ecdysone modulates Eip93 expression in late-stage egg chambers. Eip93F is in green, and DAPI is in red. A white arrowhead marks the presence of Eip93F in Dicer II (stage 11 [E¹–E³] and stage 12 [G¹–G³]) or in EcRB1^DN-overexpressing egg chambers (stage 11 [F¹–F³] and stage 12 [H¹–H³]); red arrowhead marks the absence of Eip93F in EcRB1^DN. A magenta arrowhead marks the position of nuclei in the egg chambers. (I′–J″) Eip93F function in the follicle cell mediates NC removal. Representative image of stage 14 egg chambers of control (I′ and I″) and Eip93^RNAi (J′ and J″). Egg chambers were stained with DAPI in red and Armadillo in green. (K and L) Quantification of persisting NC nuclei in stage 14 egg chambers of indicated genotypes. n indicates the number of egg chambers analyzed. The mean number of PN is provided in the graph. (K) Alternative quantification of PN. The number of PN in stage 14 egg chambers was categorized into various bins of 0 PN, 1–3 PN, 4–6 PN, 7–10 PN, 11–15 PN, and the percentage of stage 14 egg chambers in each bin was calculated (L). (M′–P″) Representative image of stage 14 egg chambers stained with Armadillo is in green and DAPI is in red. The overexpression of Eip93F in EcR^DN background rescues the clearance defect of NC nuclei in stage 14 egg chambers. A yellow arrowhead indicates PN in stage 14 egg chambers. (Q) Quantification of persisting NC nuclei in stage 14 of indicated genotypes. Error bars represent the SEM. ****P ≤ 0.0001 represent the level of significance.