Tunicamycin and thapsigargin induce ORF4 expression.

Abstract

(A) Top: Autoradiogram showing TNT of indicated plasmids. Mock: empty vector, “**”: ORF1 protein, “*”: unknown proteins. Middle: samples from top resolved by 7% SDS-PAGE, followed by autoradiography. Bottom: samples from top resolved by 15% SDS-PAGE and western using anti-ORF4. (B) Immunofluorescence of ORF4 in Huh-7 cells transfected with indicated in vitro synthesized RNA. Scale: 20μm. Shown are merged images of nuclei (blue) and ORF4 (green). “→”: positive staining, “►”: unstained. (C) Immunofluorescence of Helicase and ORF4 in Huh-7 cells transfected with in vitro synthesized wild type HEV g-1 (WT HEV) or g-3 (WT g-3 HEV) genomic RNA. Scale: 20μm. Shown are merged images of nuclei (blue) and Helicase or ORF4 (green). “→”: positive staining.</p