Figure 1.

Brefeldin A inhibits hepatitis E virus (HEV) replication. (a) Huh‐7.5 and PLC3 cells were electroporated with in vitro‐transcribed p6^SPGLuc RNA or Sar55FLuc RNA. Luciferase activities were measured at 6, 24, 48, and 72 hr post‐electroporation (p.e.). For each time point, values are presented as fold increase compared to luciferase activities measured at 6 hr p.e. (b–d) At 6 hr p.e., HEV‐p6^SPGLuc‐electroporated Huh‐7.5 cells were treated for 16 hr with brefeldin A (b and d) or sofosbuvir (c) at indicated concentrations. Luciferase activities (b and c) and viability (d) were quantified at 24, 48, and 72 hr p.e. Values are presented as a percentage of replication compared to cells treated with 0.2% dimethyl sulfoxide (DMSO). PLC3 cells electroporated with p6^SPGLuc RNA (e), and Huh‐7.5 cells electroporated with gt1 Sar55FLuc RNA (f) were treated for 16 hr with brefeldin A at indicated concentrations. Luciferase activities were measured at 24, 48, and 72 hr p.e. Values are presented as a percentage of replication compared to cells treated with 0.2% DMSO. (g) Huh‐7.5 cells were electroporated with the full‐length infectious p6 strain RNA and then treated with brefeldin A for 16 hr at indicated concentrations. At 96 hr post‐transfection, cells were fixed and analysed by immunofluorescence with an anti‐ORF2 capsid protein antibody. Values were adjusted to 100% infection for non‐treated cells (mock). Results are presented as mean ± standard deviation of three independent experiments. *, **, and *** mean p‐values below .05, .01, and .001, respectively