GSM7664017: Ad-CTRL CMC n1; Mus musculus; RNA-Seq
Mice were housed in a temperature-controlled room (22–24°C) with a 12h dark-light cycle. Animal studies were performed in compliance with European Community specifications regarding the use of laboratory animals and approved by the Nord-Pas de Calais Ethical Committee for animal use. Fat10 overexpression was performed on C57BL/6J wild-type mice (8 weeks of age) by injecting via the tail vein a solution containing 2.5x108 genome copies (GC) of a control adenovirus: Ad-CTRL or the overexpressing adenovirus: Ad-FAT10 both purchased from Genecust. Four days after adenovirus injections, mice were treated with 1 mpk of pemafibrate or vehicule (CMC) by oral gavage 18h and 5h before livers were taken. Mice were fasted 5h before the livers were taken. RNA was extracted using Trizol and reverse transcribed using the High Capacity cDNA Reverse Transcription kit (Applied Biosystems, USA). Total RNA was checked for quantity and quality using the Agilent 2100 Bioanalyzer (Agilent Biotechnologies). The library construction method and sequencing process are carried out according to the following steps: 1.Take a certain amount of total RNA samples, and use oligo dT beads to enrich mRNA with poly A tail; 2. mRNA molecules were fragmented into small pieces; 3.The fragmented mRNA was synthesized into first strand cDNA using random primers; 4. The second strand cDNA was synthesized with dUTP instead of dTTP; 5.The synthesized cDNA was subjected to end-repair and 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments; 6.Digest the U-labeled second-strand template with Uracil-DNA-Glycosylase (UDG) and perform PCR amplification; 7. Library quality control; 8. Library circularization; 9. The library was amplified to make DNA nanoball (DNB); 10. Sequencing on DNBSEQ (DNBSEQ Technology) platform.