Serpent regulates NC removal and acidification. (A ¹ –B ² ) Status of dad-lacz in clonal cells of EcRB1^DN and nonclonal control cells in stage 11 (A¹ and A²) and stage 10B (B¹ and B²). Clonal cells are marked by mCD8GFP in green. Egg chambers are stained with anti-βGAL in magenta and DAPI in cyan. Cloned cells are marked by mCD8GFP in green and encircled by a white dotted line. (C¹ and C²) Status of pMad in clonal cells of EcRB1^DN and nonclonal control cells in stage 10B. Clonal cells are marked by mCD8GFP in green. Egg chambers are stained with anti-pMad in magenta and DAPI in cyan. Clonal cells are encircled by a white dotted line. (D) Number of PN in stage 14 egg chamber of indicated genotypes as categorized into bins. The percentage of stage 14 egg chambers in each bin was calculated. n indicates the number of egg chambers analyzed. (E′–G) Status of acidified nuclei in control (E′ and E″) and in Srp^RNAi (F′ and F″). LysoTracker is in green and DAPI is in red. Yellow arrowheads mark acidified nuclei, and white arrowheads mark nonacidified nuclei. Quantification of the percentage of LysoTracker-positive NC nuclei in control and Srp^RNAi (G). n indicates the number of egg chambers analyzed and the mean percentage of LysoTracker-positive nuclei in stage 13 egg chambers of indicated genotypes. (H¹and H²) EcRB1^DN exhibits reduced levels of Srp transcripts; tub84b is the loading control. Quantification of the Srp transcripts in the EcRB1^DN-overexpressing egg chambers with respect to the control. (I¹–I⁴)srp3X-mCherry marks circulatory macrophages in magenta. Macrophages are present outside the ovarian sheath (I¹ and I²). White arrowhead marks the Srp-positive small macrophages. Schematic diagram of ovary and macrophages in magenta: top view (I³) and side view (I⁴). Error bars represent the SEM. ****P ≤ 0.0001 represents the level of significance. Source data are available for this figure: SourceData FS3.