DEV017590 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 (Adobe PDF) -

  Fig. S1. Graphs determining IC₅₀ concentrations for the COX inhibitors. The percentage of stalled egg chambers plotted against the log of the concentration of the inhibitor was used to determine a best fit line. The resulting equation from the line was used to calculate the IC₅₀ (y=50 then x=log IC₅₀).

• Supplemental Figure S2 (Adobe PDF) -

  Fig. S2. Most exogenous PGs inhibit egg chamber development in vitro. PGF_2α and PGH₂ are the only PGs tested that do not inhibit egg chamber development. PGD₂, PGE₂ and PGI₂ block egg chamber development in vitro.

• Supplemental Figure S3 (Adobe PDF) -

  Fig. S3. pxt mutants exhibit decreased pxt expression. (A-C) Whole-mount in situ hybridization using a pxt probe. (A) Expression is weak early in oogenesis and is upregulated in mid-oogenesis, S10. Expression has a punctate characteristic and the RNA has a perinuclear localization, arrows. (B) pxt^f01000 exhibits almost a complete loss of expression. (C) pxt^EY03052 egg chambers exhibit a uniform hazy expression, but lacks the punctate appearance, subcellular localization and temporal regulation. Some form of pxt being expressed in pxt^EY03052 ovaries which may restore enough Pxt function to reduce the severity of the phenotype. A-C are projections of several confocal sections.

• Supplemental Figure S4 (Adobe PDF) -

  Fig. S4. pxt mutants have altered ovariole composition. Quantification of the number of the stage indicated per ovariole of the genotype indicated normalized to the wild-type control, yw. Flies were 4 days old and grown at 25°C. pxt mutants have decreased numbers of S8 and S10 compared with wild type, and a corresponding increase in the number of S14s. Blue indicates S8; burgundy indicates S10; yellow indicates S14; n=sample size.

• Supplemental Figure S5 (Adobe PDF) -

  Fig. S5. pxt mutant ovarioles have high levels of egg chamber fusion and degeneration. Quantification of the percent of ovarioles exhibiting degrading or fused chambers in the genotypes indicated. Flies were 4 days old and grown at 25°C.

• Movie 1 -

  Movie 1. S10B egg chambers can mature in culture. Movie of time-lapse images, every 10 minutes for 10 hours, of yw S10Bs grown in culture.

• Movie 2 -

  Movie 2. COX inhibitors block egg chamber development in culture. Movie of time-lapse images, every 10 minutes for 10 hours, of yw S10Bs treated with 3 mM aspirin and grown in culture