Comparative transcriptomic responses of bank vole kidney and respiratory epithelial cells to distinct orthohantaviruses
Comparative transcriptomic responses of bank vole kidney and respiratory epithelial cells to distinct orthohantaviruses
Bank vole cell lines (MyglaSWRecB, MyglaSWTrach, MyglaAEC; Evag catalogue) immortalized with SV40 T antigen and derived from kidney, tracheal, and alveolar epithelial tissues were obtained from Bonn University. Cells were cultured at 37 °C with 5% CO₂ in DMEM supplemented with 10% heat-inactivated fetal bovine serum. PUUV Sotkamo, PHV, and Tula Moravia virus strains were provided by collaborating laboratories in Berlin and Helsinki. Viral stocks were propagated in Vero E6 cells as previously described. bank vole cells were infected with orthohantaviruses at a MOI of 0.5 and RNA extracted 5 dpi. After washing with PBS, cells were treated with Trizol (Tri Reagent, Sigma Aldrich) and chloroform. The aqueous phase containing the RNA was precipitated with isopropanol and resuspended in distilled water. Libraries were constructed using a Illumina Stranded mRNA Prep (Illumina, USA) following the supplier's recommendations. 