1. Volume 140, Issue 3, Pages 830-839.e3 (March 2011)
S-Adenosyl Methionine Improves Early Viral Responses and Interferon-Stimulated Gene Induction in Hepatitis C Nonresponders 
Jordan J. Feld, Apurva A. Modi, Ramy El–Diwany, Yaron Rotman, Emmanuel Thomas, Golo Ahlenstiel, Rachel Titerence, Christopher Koh, Vera Cherepanov, Theo Heller, Marc G. Ghany, Yoon Park, Jay H. Hoofnagle, T. Jake Liang 
Gastroenterology 
Volume 140, Issue 3, Pages e3 (March 2011)
DOI: /j.gastro
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2. Figure 1
Effects of SAMe on HCV RNA levels and on first- and second-phase kinetic responses to peginterferon and ribavirin. Viral kinetics were compared between course A (peginterferon and ribavirin alone) and course B (peginterferon, ribavirin, and SAMe), showing the effect of SAMe on (A) the first-phase viral decline, (B) the average second-phase slope, and (C) HCV RNA levels as monotherapy. P values are based on Wilcoxon matched pairs test.
Gastroenterology  , e3DOI: ( /j.gastro )
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3. Figure 2
Effects of SAMe on ISG induction in PBMCs of patients receiving peginterferon and ribavirin. Expression of (A) ISG15, (B) Mx1, and (C) RSAD2 mRNA levels in PBMCs was compared between course A (peginterferon and ribavirin alone) and course B (peginterferon, ribavirin, and SAMe) at 6 time points. Values reflect the average fold induction compared with the baseline expression [time 0] for each patient. The area under the curve reflects the mean fold induction over all time points measured. Mean AUC values were compared using the nonparametric Wilcoxon matched pairs test. (D) Protein expression of RSAD2, DMA, and actin in PBMCs was compared between baseline and 24 hours in course A, after SAMe loading at baseline, and at 24 hours in course B. STAT1 methylation was assessed by immunoprecipitation for STAT1 followed by Western blotting for DMA. Results are shown for a representative patient. Relative units (RU) were compared using time 0 for each course as baseline. *P < .05 for comparison.
Gastroenterology  , e3DOI: ( /j.gastro )
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4. Figure 3
Effects of SAMe on ISG induction in vitro. (A) Huh7.5.1 cells were treated with SAMe alone or SAMe combined with interferon alfa (50 IU/mL). Cells were incubated with SAMe for 2 hours before addition of interferon and were harvested for assessment of gene induction 4 hours after interferon treatment. ISG expression (Mx1) was evaluated by real-time PCR. (B) Huh7.5.1 cells infected with the JFH1 clone of HCV (for 24 hours) were subjected to the same treatment using SAMe alone or combined with interferon. Note the scales of Mx1 induction are different with SAMe alone and SAMe plus interferon. Mean results from 3 independent experiments are shown. *P < .05 for comparison with interferon alfa alone.
Gastroenterology  , e3DOI: ( /j.gastro )
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5. Figure 4
Effects of addition of SAMe to interferon on HCV RNA levels in vitro. Huh7.5.1 cells were infected with the JFH1 HCV for 24 hours and then treated with SAMe alone or SAMe (for 2 hours) and interferon alfa (50 IU/mL). Cells were harvested at different time points after interferon treatment: (A) 4 hrs, (B) 24 hrs, (C) 72 hrs. Intracellular HCV RNA was measured by real-time PCR. Mean results from 3 independent experiments are shown. *P < .05 for comparison with interferon alfa alone.
Gastroenterology  , e3DOI: ( /j.gastro )
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6. Figure 5
Effects of SAMe on STAT1 methylation. (A) Huh7.5.1 cells were treated with SAMe (800 nmol/L) for 2 hours, interferon alfa for 1 hour, or the combination (SAMe pretreatment followed by interferon alfa) and then harvested in RIPA buffer. Immunoprecipitation was performed with anti-STAT1, anti-PIAS1, or anti-DMA antibodies, and then Western blotting was performed with each individual antibody. The effect of SAMe treatment with or without interferon on STAT1 methylation (first panel), PIAS1 methylation (second panel), and the interaction between PIAS1 and STAT1 (third panel) is shown. (B) The experiment was repeated in the presence of HCV infection. RU, relative units; WB, Western blot.
Gastroenterology  , e3DOI: ( /j.gastro )
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7. Supplementary Figure 1
Comparison of the calculated second-phase decline in HCV RNA levels using values from weeks 1 and 2 vs from weeks 1 and 4. The second-phase slope was evaluated at 2 weeks during treatment courses A and B rather than the usual 4-week measurement for this parameter. Correlation of slopes calculated using only the first 2 weeks of course B and those calculated using values from 4 weeks are shown.
Gastroenterology  , e3DOI: ( /j.gastro )
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8. Supplementary Figure 2
Baseline expression of ISGs in PBMCs. Expression of RSAD2, ISG15, and Mx1 was compared in baseline samples determined during the same TaqMan run. Expression levels were normalized for 18S ribosomal RNA expression and arbitrarily compared with patient 1 in relative units (RU). Expression levels were compared between nonresponders (NR) and patients who became HCV RNA undetectable by PCR during treatment (PCR Neg). P values are reported from the nonparametric Mann–Whitney U test.
Gastroenterology  , e3DOI: ( /j.gastro )
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9. Supplementary Figure 3
SAMe improves ISG induction in vitro. Huh7.5.1 cells were treated with SAMe alone or 2 hours before interferon alfa therapy (50 IU/mL). Cells were harvested 4 hours after interferon treatment and (A) ISGG15 and (B) RSAD2 expression was evaluated by real-time PCR. Note the scales of Mx1 induction are different between SAMe alone and SAMe plus interferon. *P < .05 for comparison with sample treated with interferon alfa alone.
Gastroenterology  , e3DOI: ( /j.gastro )
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10. Supplementary Figure 4
SAMe improves ISG induction in primary human hepatocytes. No RSAD2 induction was seen with treatment of primary human hepatocytes with SAMe alone. SAMe treatment 2 hours before interferon alfa therapy augmented RSAD2 induction with a modest dose response similar to that seen in Huh7.5.1 cells. **P = .01 for comparison of SAMe 800 nmol/L plus interferon with interferon alone.
Gastroenterology  , e3DOI: ( /j.gastro )
Copyright © 2011 AGA Institute Terms and Conditions