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. 2016 Feb 16;113(9):E1246–E1255. doi: 10.1073/pnas.1522830113

Fig. 2.

Fig. 2.

Ced-12 and draper act in parallel pathways, and mutants show reduced fecundity. (A and B) draperRNAi Ced 12RNAi double knockdowns have a severe persisting nuclei phenotype (arrow). Stage 14 egg chambers are stained with DAPI (blue). (Scale bar, 20 μm.) (A) Control (UAS-GAL4/Sco; MKRS/TM6B) stage 14 egg chamber does not have any persisting NC nuclei. (B) Egg chamber from draperRNAi Ced-12RNAi double knockdown (UAS-GAL4/UAS-draperRNAi; GR1-GAL4 G89/UAS-Ced-12RNAi) exhibits a complete failure in NC removal with 15 persisting NC nuclei. (C) Quantification of persisting NC nuclei in stage 14 egg chambers. draperRNAi Ced-12RNAi knockdown (purple, UAS-GAL4/UAS-draperRNAi; GR1-GAL4 G89/UAS-Ced-12RNAi) has a more severe persisting nuclei phenotype than the draperRNAi knockdown (red, UAS-GAL4/UAS-draperRNAi;GR1-GAL4 G89/MKRS) or Ced-12RNAi knockdown (blue, UAS-GAL4/CyO;GR1-GAL4 G89/UAS-Ced-12RNAi). draper∆5 Ced-12RNAi double knockdown (purple, draper∆5 UAS-Ced-12RNAi/draper∆5 GR1-GAL4) has a more severe persisting NC nuclei phenotype than draper∆5 (red, draper∆5 UAS-Ced-12RNAi/draper∆5) or Ced-12RNAi (blue, draper∆5 UAS-Ced-12RNAi/GR1-GAL4 UAS-mCD8-GFP) single knockdowns. Mixed sibling controls lack either the GAL4 driver or the RNAi constructs. Data presented are mean ± SEM. ****P ≤ 0.0001. (D) Alternative quantification of data presented in Fig. 2C (see Materials and Methods). The draper Ced-12 double knockdowns result in a stronger persisting nuclei phenotype than draper or Ced-12 single knockdowns. (E and F) Intact ovaries stained with DAPI (blue). (Scale bar, 100 μm.) (E) Control [FRT 2A/Df(3L)BSC181] ovary does not contain lingering NC nuclei. The center of the ovary contains FCs that have been shed from mature stage 14 egg chambers as they enter the oviduct (arrows). (F) NC nuclei accumulate in homozygous draper∆5 ovary (arrows). (G) draperRNAi Ced-12RNAi double knockdowns in stretch FCs (PG150-GAL4/+; UAS-GAL4/UAS-draperRNAi; +/UAS- Ced-12RNAi) have reduced egg laying compared with the control (PG150-GAL4/+; UAS-GAL4/+; UAS-luciferaseRNAi/+). Egg laying was quantified in females 21–25 d old. n, number of females. Data presented are average number of eggs laid per female per day ± SEM. **P ≤ 0.01.