biostudies-arrayexpressUnknownTranscriptomicsGenomicsProteomicsNares TrakooljulNextSeq 2000RNA-seq of coding RNADrosophila melanogasterDrosophila melanogasterhttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15429This experiment aims to understand how chronic dietary reserpine treatment affects gene expression in response to acute heat shock in young (12-day-old +24hr heat stress) and aged (43-day-old) male Drosophila melanogaster in the context of aging. Canton S flies were chronically fed either a reserpine-supplemented (1000µM) or control diet (DMSO) starting from eclosion (1 day old). The whole flies were snap frozen and subjected to RNA-seq analysis at age of 13 days under the heat stress condition and at age of 43 days under the standard condition.biostudies-arrayexpressSample Treatment - Reserpine (Sigma-Aldrich, Cat. No. 83580; MW 608.68 g/mol) was dissolved in dimethyl sulfoxide (DMSO) to generate a 100 mM stock solution (760 mg in 12.5 mL DMSO). After autoclaving and cooling the fly food to approximately 60°C, reserpine was added to achieve final concentrations of 0 (DMSO control) or 1000 μM. Fly food was prepared with a standardized cornmeal–soy–molasses medium (containing 3.21 L distilled water, 30 g soy flour, 260 g corn flour, 60 g active dry yeast, 26 g agar, 260 g molasses, and 130 g malt extract per batch), homogenized and autoclaved at 121°C for 20 minutes. After cooling to ~60°C, 83 mL of 10% (w/v) methylparaben (in ethanol) and 59 mL acid mix (60 mL 85% phosphoric acid, 418 mL propionic acid, distilled water to 2 L) were added, and the medium was dispensed aseptically (8 mL per vial) and allowed to solidify.Growth Protocol - Drosophila melanogaster (Canton-S wild type) were reared at 25°C under a 12:12 hour light: dark cycle. For the heat stress condition, 12-day old flies were exposed to 24 hours of heat shock at 31°C.Nucleic Acid Extraction - Flies were flash-frozen in liquid nitrogen, and total RNA was extracted using the Monarch Total RNA Miniprep Kit (NEB). RNA purity and integrity were assessed by NanoDrop (A260/A280 ~2.0) and Agilent Bioanalyzer.Sample Collection - Flies were collected at age of 13 days (12 days at 25°C followed by 24 hours at 31°C) for the heat stress condition and at age of 43 days for the normal condition. Flies were flash-frozen in liquid nitrogen and stored at -80°C until RNA extraction.Sequencing - DNA libraries were multiplexed and paired-end sequenced for 2x101bp reads at 750 pM concentration on the NextSeq 1000/2000 system using a P3 flowcell at the sequencing facility of Research Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.Library Construction - One microgram of total RNA of was used for library preparation using an Illumina Stranded mRNA Prep Ligation kit according to the manufacture's recommendation (Illumina). The libraries were quality checked for fragment length distribution on Agilent Bionalyzer and normalized to an equal concentration of 10nM prior to pooling.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesNares TrakooljulShahaf PelegfalseRNA-Seq of male Drosophila chronically treated with dietary reserpine under control and heat stress conditionsThis experiment aims to understand how chronic dietary reserpine treatment affects gene expression in response to acute heat shock in young (12-day-old +24hr heat stress) and aged (43-day-old) male Drosophila melanogaster in the context of aging. Canton S flies were chronically fed either a reserpine-supplemented (1000µM) or control diet (DMSO) starting from eclosion (1 day old). The whole flies were snap frozen and subjected to RNA-seq analysis at age of 13 days under the heat stress condition and at age of 43 days under the standard condition.2026-05-04T00:00:00Z2026-05-04T06:57:45.03Z2025-07-31T22:57:34.042ZE-MTAB-15429ERP177676EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003738EFO_0004184EFO_0003969