Fig. 5

CRM1 is directly involved in HBV virion secretion. A In an HBV transient transfection system in HuH-7 cells, Lentivirus shCRM1 #1 and #2 significantly knocked down CRM1 expression and reduced HBV virion secretion. The bar graph (right panel) is based on the signal intensities of virions and naked capsids (left panel) using densitometry and Image J. The mean values were calculated from at least three independent experiments. shScramble: a non-specific control. The vertical dotted line indicates splicing of data from the same gel. HBc protein signals of secreted virions (solid bar) and naked capsids (open bar) in the shScramble experiment, are shown here as a 100% reference control. Both CRM1 and HBc proteins in the total intracellular lysates were assayed by SDS-PAGE. B The reduction of HBV virion secretion by 0.1 μM SINE compound treatment can be rescued by cotransfection with a drug-resistant mutant CRM1 expression vector (see Methods). The bar graph (right panel) is based on the signal intensities of virions and naked capsids (left panel) using densitometry and Image J. The mean values were calculated from at least three independent experiments. The vertical dotted lines indicate splicing of data from the same gel. HBc protein signals of secreted virions (solid bar) and naked capsids (open bar) in the no treatment experiment (HBV WT only), are shown here as a reference control (1.0)