SEARCH

SEARCH BY CITATION

This article includes Supplementary Figures available at http://www.interscience.wiley.com/jpages/0270-9139/suppmat

FilenameFormatSizeDescription
suppmat_1295_fig1.tif91KExperimental design for cell culture and the corresponding treatments. Hepatocytes were isolated by collagenase perfusion and cultured with William's medium E containing 5% FBS for two hours. Afterwards, the medium was replaced by fresh medium without FBS and the culture was maintained for 18 hours. After the removal of culture medium, L-NAME or PGE1 were added at 4 or 2 hours, before D-GalN, respectively. Hepatocytes were harvested at the selected time points after PGE1 or D-GalN treatment.
suppmat_1295_fig2.tif115KEffect of PGE1 on cytokine-induced NOS-2 mRNA expression in the presence or absence of D-GalN in cultured hepatocytes. PGE1 (1 mol/L) was administered 2 hours before cytokines (200 U/mL IL-1 and 200 U/mL IFN-g) or D-GalN (5 mmol/L) in primary cultures of rat hepatocytes. The measurement of NOS-2 mRNA expression was carried out 12 hours after treatments by RT-PCR. Two bands of 216 bp and 138 bp corresponding to b-actin and NOS-2 mRNA, respectively, were observed in agarose gels. The blots are representative of two independent experiments.
suppmat_1295_fig3.tif151KProposed mechanism of negative inhibition of NOS-2 gene expression by PGE1-induced NO production. Pre-administration of PGE1 to hepatocytes activates NF-kB translocation resulting in NOS-2 expression, mRNA synthesis, NOS-2 protein translation and NO synthesis from L-arginine. NO synthesized exerts a negative feedback regulation on NF-kB activation and further NOS-2 expression induced by D-GalN and/or cytokines.

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.