biostudies-arrayexpressUnknownTranscriptomicsGenomicsProteomicsFabian GrammesNextSeq 1000RNA-seq of coding RNASalmo salarSalmo salarhttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16379In vitro challenge of primary hepatocytes, with the Infectious pancreatic necrosis (IPN) virus. Cells from were infected at MOI 0.1 with two different isolates of IPNV: V1244 and Vir410/2018. V1244 represents classical IPNV that is affected by the IPN-QTL, while Vir410/2018 is the QTL-insensitive or recent isolate, rIPNV in short.biostudies-arrayexpressGrowth Protocol - Liver cells were cultured at 20°C in hepatocyte culture media for a period of two days. After that time, the cells were washed with PBS and fresh culture media was added supplemented additionally with growth factors (5 ng/mL epidermal growth factor (EGF) and 3 ng/mL hepatocyte growth factor (HGF)). At day 7 post-culture, the media was replaced once again to hepatocyte culture media without growth factors.Sequencing - Sequencing was conducted by NovoGene and BmkGeneLibrary Construction - Libraries were constructed by NovoGene and BmkGeneSample Collection - To establish a primary hepatocyte culture, 200 g fish from the Fish Centre facility at NMBU were euthanized by a sharp blow to the head, and the hepatocytes were isolated as previously described: Datsomor, A.K.; Wilberg, R.; Torgersen, J.S.; Sandve, S.R.; Harvey, T.N. Efficient Transfection of Atlantic Salmon Primary Hepatocyte Cells for Functional Assays and Gene Editing. G3 Genes|Genomes|Genetics 2023Nucleic Acid Extraction - total RNA was extracted using the RNeasy Mini kit (Qiagen®, following the manufacturer’s protocol for extraction from cells. he RNA samples were labelled and stored at -80°C until processed downstream.Sample Treatment - Two independent challenge tests using primary hepatocytes from 4 different fish (F1 – F4) were performed. Challenge 1 consisted of cells from F1 while the hepatocytes from the remaining fish (F2 – F4) were part of challenge 2. The cells from both challenges were infected in duplicates at MOI 0.1 with cIPNV or rIPNV for 1 hour at 15°C. After incubation time, the viral inoculum was removed and replaced with infection media containing also Amphotericin B (2.5 µg/mL). The cells were kept at 15°C and monitored daily for CPE. Both cells and supernatants were collected for RNA extraction at 4 and 7 dpi for challenge 1 and at 2 and 4 dpi for challenge 2.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesFabian GrammesfalseIPN in Vitro ChallengeIn vitro challenge of primary hepatocytes, with the Infectious pancreatic necrosis (IPN) virus. Cells from were infected at MOI 0.1 with two different isolates of IPNV: V1244 and Vir410/2018. V1244 represents classical IPNV that is affected by the IPN-QTL, while Vir410/2018 is the QTL-insensitive or recent isolate, rIPNV in short.2025-12-24T00:00:00Z2025-12-24T02:01:28.567Z2025-12-11T12:58:58.648ZE-MTAB-16379ERP186427EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003738EFO_0004184EFO_0003969