Three-month-old adult male zebrafish were used for the VHSV challenge experiments. The fish were adapted to the experimental temperature described in the mortality experiment and injected with 1 × 105 TCID50/mL of VHSV. After the initial injection, the VHSV copy number was checked at 12 h postinfection (hpi) to evaluate the injection process.
Growth protocol
Wild-type (WT) and viperin-/- zebrafish were maintained as previously described by Avdesh at el. Water used in tanks was maintained at constant pH (pH 6.8–7.5), conductivity (500–800 mS), and temperature (28°C ± 0.5°C) during the fish rearing. The zebrafish facility’s light and dark cycle consisted of 14:10 h. The fish were fed three times with the Artemia diet. The Jeju National University Animal Ethics Committee reviewed and approved this study
Extracted molecule
total RNA
Extraction protocol
TRIzol reagent (Thermo Fisher Scientific, Waltham, MA, USA) was used for RNA extraction.
Label
SYBR Green
Label protocol
All qPCR reactions were performed in triplicate in a reaction volume of 10 μL containing 4 μL of cDNA template, 0.4 μL of each qPCR primer (10 pmol/μL), 5.0 μL of 2× TB Green® Premix Ex Taq™ II (TaKaRa, Japan), and 1.2 μL of PCR-grade water. Reactions were run in a Real-Time system Thermal Cycler Dice™ system III TP950 (TaKaRa, Japan) under the following conditions: 95 °C for 10 s, followed by 45 cycles at 95 °C for 5 s, 60 °C for 10 s, and 72 °C for 20 s, and a final cycle at 95 °C for 5 s, 60 °C for 30 s, and 95 °C for 15 s.
Hybridization protocol
n/a
Scan protocol
n/a
Description
Test SAMPLE 19
Data processing
The qPCR primers for gene expression analysis were designed according to the guidelines for the publication of quantitative realtime PCR experiments (MIQE) experiments Fold Change worksheet reports test/control. The respective test value was divided with the respective control value.
