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

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

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

Reactivation of Hepatitis B Virus in HBsAg-Negative Patients with Hepatocellular Carcinoma - Fig 2

<p>A, Kaplan-Meier curves of HBV reactivation for the HCC and control groups. Patients with HCC undergoing TACE had significantly more frequent episodes of HBV reactivation during follow-up than controls without TACE, with estimated 12- and 24-month rates of 7.7% vs. 0% and 15.9% vs. 2.0%, respectively (<i>P</i> = 0.006, log-rank test). B, Comparison of HBV reactivation between the HCC and control groups. The risk of HBV reactivation was highest with combo + RT, followed by combo-TACE and mono-TACE, with estimated 24-month rates of 44.3%, 37.8%, and 6.2%, respectively (<i>P</i> = 0.001, log-rank test). C, Comparison of HBV reactivation according to prior CHB status. Patients with prior CHB had a significantly higher incidence of HBV reactivation than those without, with estimated 24-month rates of 53.7% and 7.5%, respectively (<i>P</i> < 0.001, log-rank test).</p

Clinical features of 13 HBsAg-negative patients who developed HBV reactivation during the follow-up.

<p>HBsAg, hepatitis B surface antigen; HBV, hepatitis B virus; Pt, patient; M, male; F, female; TACE, transarterial chemoembolization; Mono, mono-drug TACE; Combo, combination-drug TACE; RT, radiotherapy; mo, months; UD, undetectable; No, number; Tx, treatment; ALT, alanine aminotransferase; SDH, subdural hemorrhage; PD, progressive disease; FU, follow-up.</p><p>Clinical features of 13 HBsAg-negative patients who developed HBV reactivation during the follow-up.</p

Risk factors for HBV reactivation in patients with HCC receiving transarterial therapy.

<p>HBV, hepatitis B virus; HCC, hepatocellular carcinoma; No, number; HR, hazard ratio; CI, confidence interval; ALT, alanine aminotransferase; HCV, hepatitis C virus; TACE, transarterial chemoembolization; RT, radiotherapy.</p><p>*<i>P</i> for trend.</p><p>Risk factors for HBV reactivation in patients with HCC receiving transarterial therapy.</p

Patient enrollment and assessment.

<p>Patient enrollment and assessment.</p

A and B, Clinical hepatitis associated with HBV reactivation according to treatment intensity.

<p>(A) ALT levels at HBV reactivation in reactivated patients (<i>P</i> = 0.321). (B) Hepatitis due to HBV reactivation according to treatment intensity among the entire patient group with HCC. The incidence rates of reactivation hepatitis were 1.3% (1/75) for mono-TACE, 5.0% (1/20) for combo-TACE, and 14.3% (2/14) for combo + RT (<i>P</i> for trend = 0.021).</p

Baseline characteristics of the study subjects.

<p>Treatment group = HBsAg-negative patients with HCC undergoing anti-cancer therapy (n = 109).</p><p>Control group = HBsAg-negative patients with cirrhosis (n = 16) and patients with HBsAg seroclearance (n = 46).</p><p>ALT, alanine aminotransferase.</p><p>Baseline characteristics of the study subjects.</p