Project description:The outcome of Notch activation of proliferation depends on cellular context. In Drosophila wing discs Notch pathway overactivation results in hyperplasia. To understand the mechanisms we have used genomic strategies to indetify the Notch-S(H) target genes directly regulated in wing disc hyperplasia. These data are the results from expression profiling the RNAs from hyperplastic wing discs overexpressing Su(H). Direct comparison of Giant third instar lavae wing imaginal disc (UAS-GFP:Su(H) expressed by the patched[559.1]-Gal4 driver) vs control (UAS-NLS-GFP expressed by the patched[559.1]-Gal4 driver). 3 Biological replicates, the 3rd replicate was performed as a dye-swap.
Project description:The outcome of Notch proliferation on proliferation depends on the context. In Drosophila wing imaginal discs Notch activation causes hyperplasia despite having localized inhibitory effects on proliferation. To understand te underlying mechanisms we have used genomic strategies to identify the Notch-Su(H) target genes during wing discs hyperplasis. these data are the results from expression profiling the RNAs from hyperplastic wing discs overexpressing Nicd. Direct comparison of third instar lavae wing imaginal disc Nicd (abxUbxFLPase; Act>y>Gal4, UAS GFP; FRT82B tubGal80 with UAS-Nicd; FRT82B) vs control (abxUbxFLPase; Act>y>Gal4, UAS GFP; FRT82B tubGal80 with FRT82B ). 4 Biological replicates, the 2nd replicate was performed as a dye-swap.
Project description:The strongest risk factor for developing Alzheimer's Disease (AD) is age. Here, we study the relationship between ageing and AD using a systems biology approach that employs a Drosophila (fruitfly) model of AD in which the flies overexpress the human AM-NM-242 peptide. We identified 712 genes that are differentially expressed between control and AM-NM-2-expressing flies. We further divided these genes according to how they change over the animal's lifetime and discovered that the AD-related gene expression signature is age- independent. We have identified a number of differentially expressed pathways that are likely to play an important role in the disease, including oxidative stress and innate immunity. In particular, we uncovered two new modifiers of the AM-NM-2 phenotype, namely Sod3 and PGRP-SC1b. Transcript level measured using microarrays in biological quadruplicate (except day 20 which is in biological triplicate), each array is AM-NM-2 vs control at the same timepoint (defined by % survival), one replicate per array with dye-swaps.
Project description:Dichaete is a developmentally important transcription factor, known to be involved in basic biological processes including segmentation and nervous system development among others. The aim of this experiment was to gain further insight into the role of Dichaete during early embryogenesis, specifically looking at targets in the midline using dominant negative constructs expressed via the UAS/Gal4 system, using simGal4 to drive expression in the midline. 4 independent biological replicates. 3.5-4.5h old embryos were collected, and the RNA was extracted using Trizol. The UAS-dominant negative construct expressing embryos were compared to UAS-GFP expressing controls. Construct expression was driven using simgal4.
Project description:Dichaete is a developmentally important transcription factor, known to be involved in basic biological processes including segmentation and nervous system development among others. The aim of this experiment was to gain further insight into the role of Dichaete during early embryogenesis, by looking at the disruption of gene expression in Dichaete mutants. Stage 10-11 embryos (5 and 7.5 hours after egg laying) from a cross between Dr72/TM3, twi-GAL4 UAS-Gfp Dr513/TM3, twi-GAL4 UAS-Gfp, were hand picked under a fluorescence dissecting microscope. GFP negative homozygous Dichaete mutant embryos and their heterozygous single GFP positive siblings were collected and approximately 150 embryos per sample were stored frozen in Trizol
Project description:Dichaete is a developmentally important transcription factor, known to be involved in basic biological processes including segmentation and nervous system development among others. The aim of this experiment was to gain further insight into the role of Dichaete during early embryogenesis, specifically looking at targets in the midline using dominant negative constructs expressed via the UAS/Gal4 system, using prosGal4 to drive expression in developing neuroblasts. 4 independent biological replicates. 3.5-4.5h old embryos were collected, and the RNA was extracted using Trizol. The UAS-dominant negative construct expressing embryos were compared to UAS-GFP expressing controls. Construct expression was driven using prosGal4.
Project description:Timecourse of gene expression changes in Drosophila SoxN homozygous mutant embryos compared with their heterozygous siblings, from stage 7 to 13 of embryonic development. Five time points: stage 7-8, stage 9, stage 10, stage 11 and stage 12-13. Four biological replicates per time point. Two conditions: SoxN homoxygous vs SoxN heterozygous mutant embryos.
Project description:Genome-wide binding of SoxN in wild type embryos: SoxNDam (DamID, stage 8-11), SoxND1 (ChIP, stage 8-11), SoxND2 (ChIP, stage 8-11), SoxNPA179 Early (ChIP, stage 7-10), and SoxNPA179 Late (ChIP, stage 11-13). One DamID experiment, four ChIP experiments. Three biological replicates per experiment. Two conditions: Dam (control) vs SoxNDam (sample) for DamID, anti-M-NM-2gal (control, 40-1a antibody from DSHB) vs anti-SoxN (sample, either SoxND1, SoxND2 or SoxNPA179 antibody) for ChIP. The only commercially available antibody is the anti-BetaGal. All the anti-SoxN antibodies are home-made. anti-BetaGal: Developmental Studies Hybridoma Bank (DSHB), 40-1a, mouse, monoclonal (http://dshb.biology.uiowa.edu/beta-galactosidase_2) SoxND1: polyclonal antiserum raised in rabbit immunized with a protein fragment corresponding to amino acids 2-92 of SoxN. Produced by the modENCODE consortium (http://www.modencode.org/). SoxND2: polyclonal antiserum raised in rabbit immunized with a protein fragment corresponding to amino acids 317-417 of SoxN. Produced by the modENCODE consortium (http://www.modencode.org/). SoxNPA179: affinity purified polyclonal antibody raised in rabbit immunized with a small peptide corresponding to amino acids 506-521 of SoxN. Commisioned by our lab and produced by Eurogentec (http://www.eurogentec.com/eu-home.html).