Journal: G3: Genes|Genomes|Genetics

Article Title: Defects in the Neuroendocrine Axis Contribute to Global Development Delay in a Drosophila Model of NGLY1 Deficiency

doi: 10.1534/g3.118.300578

Figure Lengend Snippet: Positive correlation between 3X replicates. A) The three pairwise comparisons of Z-scores show positive correlations indicating that a set of small molecules could modify the small larval size phenotype B) The positive correlations between replicates is more evident when only plotting Z-scores of < -2 or > 2. C) Larval size is partially rescued when 20E is fed to Pngl PL homozygous larvae.

Article Snippet: 20E (Enzo) was dissolved in 100% ethanol and added to molten standard fly food media at 200μM and added to vials.

Techniques:

Journal: G3: Genes|Genomes|Genetics

Article Title: Defects in the Neuroendocrine Axis Contribute to Global Development Delay in a Drosophila Model of NGLY1 Deficiency

doi: 10.1534/g3.118.300578

Figure Lengend Snippet: 20-hydroxyecdysone (20E), but not an earlier ecdysone pathway precursor (7-dehydrocholesterol), partially rescues developmental delay and lethality in Pngl PL mutants . The time to pupation of A) Pngl PL/+ heterozygous or B) Pngl PL homozygous larvae reared on food treated with different concentration of 20E (n = 10 replicates, 20-30 individuals/replicate). 20E did not impact developmental rate of heterozygous Pngl PL larvae to pupation, but partially rescued development delay of homozygous Pngl PL larvae to pupation at 200µM. The time to pupation of C) Pngl PL/+ heterozygous or D) Pngl PL homozygous larvae reared on food treated with different concentration of 7-d (n = 3 replicates, 25-30 individuals/replicate). 7-d did not impact developmental rate of heterozygous or homozygous Pngl PL larvae to pupation. E) The fraction of animals surviving to eclosion. 20E, but not 7-d, rescued larval lethality of Pngl PL homozygous at 200µM. (n = 10 replicates, 20-30 individuals/replicate; n = 3 replicates, 25-30 individuals/replicate, respectively).

Article Snippet: 20E (Enzo) was dissolved in 100% ethanol and added to molten standard fly food media at 200μM and added to vials.

Techniques: Concentration Assay

Journal: G3: Genes|Genomes|Genetics

Article Title: Defects in the Neuroendocrine Axis Contribute to Global Development Delay in a Drosophila Model of NGLY1 Deficiency

doi: 10.1534/g3.118.300578

Figure Lengend Snippet: Pngl larvae are ≥10X more sensitive to the proteasome inhibitor bortezomib. Images A) and quantification of larval size B) of 3 day Pngl PL/+ heterozygous or age matched Pngl PL/PL mutant larvae raised on food treated with 0, 1, 5, 10 or 25µM bortezomib. Black points report the size (in arbitrary units) of individual larvae. Blue lines report the mean and SE of these populations. Bortezomib delays larval developmental progression in Pngl PL homozygous larvae more severely than Pngl PL/+ heterozygous larvae leading to smaller sized larvae, and is lethal for homozygotes at concentrations equal to or greater than 5 µM. For these experiments, Bortezomib was solubilized in DMSO, resulting in a final concentration of DMSO in treated food of 0.025%, which is below the threshold of effect on Pngl PL homozygous larvae. (** significant at P < 0.01; Student’s t -test) C) Quantification of larval size for Pngl PL homozygous larvae raised on untreated food, or food treated with 0.1% DMSO, 1 µM bortezomib, 0.1% DMSO and 1 µM bortezomib, or 0.1% DMSO, 1 µM bortezomib, and 133 µM 20E. 20E partially rescued the effect of bortezomib treatment on larval size in Pngl PL homozygotes. (** significant at P < 0.01; Student’s t -test).

Article Snippet: 20E (Enzo) was dissolved in 100% ethanol and added to molten standard fly food media at 200μM and added to vials.

Techniques: Mutagenesis, Concentration Assay