Fig. 2: In-vitro characterization of HVR sequences identified by clonal sequencing.

The identified insertions were cloned into the HEV reporter or full genome of the Kernow-C1-p6 strain, thereby replacing the HVR with insertion containing HVRs as indicated in Fig. 1. a Replication kinetics of HVR constructs with Kernow-C1-p6 (p6) and Kernow-C1-p1 (p1) as references are shown. Plotted is the time post electroporation as well as mean (+/- SD) relative light units (RLU) normalized to the four-hour value of n = 7 biologically independent experiments for p1 h.TRIM22, n = 3 for dup constructs and n = 6 for other constructs. b The HEV replicon system was used to analyse the ribavirin (RBV) sensitivity by treating the cells for five days post-electroporation with RBV concentrations ranging from 0.19 µM to 100 µM. Plotted is the mean (+/- SD) HEV replication as a percentage of untreated controls in n = 3 biologically independent experiments for dup constructs, n = 6 for other constructs. Lines represent dose-response curves of four-parameter log-logistic analysis. c The full-length system was used to produce infectious particles, which were titrated onto HepG2/C3A cells to determine the achieved viral titers as FFU/mL via immunofluorescence. A representative picture of a whole 96-well infected with non-enveloped HEVcc stained for the ORF2 protein (black) is shown above each column. Plotted are the means with standard deviation (+/- SD) of n = 8 biologically independent experiments for p1 and p6, n = 3 for other constructs. Source data are provided in the Source Data file.