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. Author manuscript; available in PMC: 2011 Jul 22.

Published in final edited form as: Hepatology. 2010 Jul;52(1):47–59. doi: 10.1002/hep.23671

Fig. 8 — (A) Huh7 cells transfected with JFH1 RNA were cultured with heat-inactivated serum overnight, serum-starved for 24 hrs, and treated with indicated concentrations of recombinant human TGFβ1 (R&D Systems) daily for another 72 hrs. Then, samples were analyzed for Nox4 mRNA by qPCR. Data were normalized by GAPDH mRNA content. (B) Cell culture medium from control and JFH1 cells were analyzed for TGFβ1 concentration. Data were normalized by cell number and expressed as percentage of controls, where control values were 2,133.5 ± 277.9 and 2,720.8 ± 359.6 pg per 10⁶ cells at 72 hrs and 43 days, respectively. TGFβ1 present in the cell culture serum was subtracted from the readings. (C) Control and JFH1 cells were incubated with 2 μg/ml of anti-TGFβ1 antibodies (R&D Systems) and, after 48 or 72 hrs, analyzed for Nox4 protein concentration by Western blots. Nox4 protein was quantified by densitometry and normalized by the level of β-actin. * indicates statistically significant difference from controls (P < 0.05). (D) Proposed role of hepatocyte Nox proteins in HCV-induced pathogenesis.

Fig. 8 — (A) Huh7 cells transfected with JFH1 RNA were cultured with heat-inactivated serum overnight, serum-starved for 24 hrs, and treated with indicated concentrations of recombinant human TGFβ1 (R&D Systems) daily for another 72 hrs. Then, samples were analyzed for Nox4 mRNA by qPCR. Data were normalized by GAPDH mRNA content. (B) Cell culture medium from control and JFH1 cells were analyzed for TGFβ1 concentration. Data were normalized by cell number and expressed as percentage of controls, where control values were 2,133.5 ± 277.9 and 2,720.8 ± 359.6 pg per 10⁶ cells at 72 hrs and 43 days, respectively. TGFβ1 present in the cell culture serum was subtracted from the readings. (C) Control and JFH1 cells were incubated with 2 μg/ml of anti-TGFβ1 antibodies (R&D Systems) and, after 48 or 72 hrs, analyzed for Nox4 protein concentration by Western blots. Nox4 protein was quantified by densitometry and normalized by the level of β-actin. * indicates statistically significant difference from controls (P < 0.05). (D) Proposed role of hepatocyte Nox proteins in HCV-induced pathogenesis.