FIG 7.

HA-tagged ORF1 protein is membrane-associated. Hep293TT cells electroporated with full-length B2HA or parental (wild type [wt]) HEV RNA were lysed in a hypotonic buffer, followed by membrane flotation as described in Materials and Methods. Lysates were not treated (−) or treated with 1% Triton X-100 (Tx) prior to gradient centrifugation. Eight fractions were collected from the top and analyzed by immunoblotting using rabbit MAb C29F4 against the HA tag. Detection of CLIMP63 served as a control for an integral membrane protein.