Fig. 6: Characterization of generated artificial insertions in the HEV HVR.

a Two types of artificial insertions were created and cloned in the Kernow-C1-p1 (p1) and Kernow-C1-p6 (p6) reporter replicon. A rigid XP linker, mimicking the high proline content of the HVR and one with a more flexible linker. Additionally, NLS sequences (IPMK, IP3KB, SHIP1, SV40) were incorporated in the middle of the artificial insertions. See Supplementary Movies 14–17. b HEV replication kinetics were measured with p1 and p6 as reference (both grey) while constructs of interest are depicted in purple shades. Plotted are the mean (+/- SD) relative light units (RLU) normalized to the four-hour value over time (hours post electroporation) of n = 3 biologically independent experiments. c For comparison the replication values 96 h p.t. of all constructs are plotted as column diagram.+/- SD d Huh7 cells were transfected with plasmids encoding a triple eYFP in tandem with the artificial insertions including p1 and p6 flanking regions, respectively. Cells were fixed after 16–20 hours and the mean fluorescence intensity (MFI) for eYFP was measured for each compartment. Shown are example cells in 3D. eYFP is shown in green, the cell surface is depicted in white while the nuclear surface is depicted in red. See Supplementary Movies 18–25. e The eYFP MFI was measured for each compartment for ten n = 11 cells per construct, and individual outliers were removed by applying ROUT (Q = 1) method implemented in GraphPad Prism. Depicted is the range of individual data points as violin plots with a straight line as the median and dashed lines as quartiles. Source data are provided in the Source Data file.