characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-05-08T17:00:43Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:49:40Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2223465
characteristics.characteristic: External Id
characteristics.text: College of Life Science and Technology, Tongji University, Shanghai, China
characteristics.characteristic: INSDC center name
characteristics.text: 2014-05-08T17:00:43Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:49:40Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS359851
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1966:GFP37
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: fruit fly
characteristics.characteristic: common name
characteristics.text: Protocols: Stocks used are UAS-mCD8GFP, UAS-Brm-IR (#31712), and Hs-Gal4 from Bloomington Stock Center (http://flystocks.bio.indiana.edu). All transgenes are in the third chromosome. We crossed UAS-Brm-IR males with Hs-Gal4 females to generate the transgenic strain containing UAS-Brm-IR and Hs-Gal4 for Brm knockdown (experimental group, short for BrmIR). Similarly, the transgenic strain containing UAS-mCD8GFP and Hs-Gal4 (control group, short for GFP) was generated by crossing UAS-mCD8GFP males with Hs-Gal4 females and expected to remove the influence of UAS-Gal4 system. Both control and experimental group flies were cultured at normal temperature (25C) for 83.5h (i.e., the third larvae stage) after fertilization. Next, they were cultured at 37C for 30 min to knockdown Brm by the expression of Brm shRNA through heatshock in waterbath, and followed recovery at 25C for 4.5hr. Repeat the heatshock-and-recovery treatment twice, but with recovery time of 3.5hr and 2hr, respectively. Then, the larvae were collected and used in this study. These two groups of flies were denoted as GFP37 and BrmIR37, respectively. Meanwhile, the two groups of flies were cultured at 25C all the time in parallel after fertilization as negative controls. These flies were denoted as GFP25 and BrmIR25, respectively. Total RNA was extracted from 95-99h larvae using TRIzol (Invitrogen). Reverse transcription was performed on 500ng of total RNA in 10uL reaction system for each sample using PrimeScript RT Master Mix Perfect Real Time Kit (TAKARA, Code: DRR036A). The Realtime PCR was carried out in triplicate using F-416L DyNAmoTM ColorFlash SYBR Green qPCR Kit (Thermo). The RNA sequencing libraries were constructed from the extracted RNA using standard Illumina libraries prep protocols.
characteristics.characteristic: description
characteristics.text: larval stage
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000069
characteristics.characteristic: developmental stage
characteristics.text: control
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001461
characteristics.characteristic: genotype
characteristics.text: Drosophila melanogaster
characteristics.characteristic: organism
characteristics.text: Drosophila melanogaster
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_7227
characteristics.characteristic: organism
characteristics.text: Drosophila melanogaster
characteristics.characteristic: scientific_name
characteristics.text: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: GFP37
characteristics.characteristic: title