DUSP1 suppression increases STAT1 activity and nuclear translocation.

<p>Phosphorylation (A) and nuclear translocation (B) of STAT1 were assessed by cell-based ELISA and confocal microscopy, respectively. (A) Expression of phospho-STAT1 was normalized to that of STAT1 and then to the ratio in LV-cont-infected cells. (B) Red, anti-STAT1; blue, DAPI. Merged image allows assessment of nuclear localization of STAT1; original magnification ×200. All data are representative of at least three independent experiments.</p

Synergistic effects of IFN-α with DUSP1 depletion against HCV.

<p>Cells were treated with 0–10⁴ IU/mL IFN-α for 72 h. HCV RNA was measured by rqRT-PCR and normalized to β-actin and then to LV-shDUSP1-infected cells not treated with IFN. All data are representative of at least three independent experiments. **<i>P</i> < 0.01.</p

Overexpression of DUSP1 rescues anti-HCV effect by DUSP1 silencing.

<p>LV-shDUSP1 cells were transfected with pcDNA3.1-DUSP1 plasmid. (A) At 72 h post-transfection, the level of HCV proteins were determined by western blot analysis. (B) Localization of STAT1 was detected using immunocytochemistry. Red, anti-STAT1; blue, DAPI. White narrows indicate cytoplasm location of STAT1; original magnification ×200. All data are representative of at least three independent experiments.</p

DUSP1 silencing has an antiviral effect in the FK replicon and HCV cc system.

<p>(A) HCV RNA was measured in LV-shDUSP1-infected cells by rq-RT-PCR and normalized to β-actin and expressed relative to levels in LV-cont-infected cells. (B) HCV NS5A, HCV NS5B, DUSP1, and β-actin protein were measured by Western blot analysis and normalized to β-actin. (C and D) Direct effects of DUSP1 knockdown were confirmed by transfection of DUSP1-siRNA in HCVcc system. At 72 h post-transfection, HCV (C) and DUSP1 mRNA (D) were measured by rqRT-PCR and normalized to β-actin and G6PD, respectively, then to expression in control cells. All data are representative of at least three independent experiments. *<i>P</i> < 0.05; **<i>P</i> < 0.01; ***<i>P</i> < 0.001 compared to the control. si scram, scrambled siRNA; si DUSP1, DUSP1 siRNA.</p