Stat5 regulates Tmprss6 expression and this participates to the regulation of hepcidin expression by IL-6 treatment.

<p>(A) Hepa 1–6 cells were transfected with mouse wild-type or mutant <i>Tmprss6</i> promoter-luciferase construct, in presence of empty vector or Stat5b plasmid. After 48 hours, luciferase activity was determined. The assay was performed in duplicate for a total of 6 independent experiments. For each experiment, raw data were normalized to the value of luciferase activity in cells transfected with the empty vector. Values shown are mean of normalized luciferase activities+/− SEM. Mean in treated groups was compared to 1 by one sample student t tests. (B) Hep3B cells were transfected with siRNA control or siRNA Stat5b and analyzed for <i>TMPRSS6</i> relative to <i>RPLl19</i> mRNA expression. For each experiment, raw data were normalized to the expression value in mock-treated cells. Values shown are means of normalized expression values obtained in 3 independent experiments+/− SEM. Means in siRNA control and siRNA STAT5b groups were compared by student t tests. (C) Hep3B cells were transfected with siRNA control or siRNA TMPRSS6 and treated without or with 20 ng/mL of IL-6, followed by analysis of <i>HAMP</i> relative to <i>RPL19</i> mRNA expression. The mean 4 independent experiments is presented. Values shown are means of expression values divided by a calibrator quantity (the mean value of expression for the cells transfected with siRNA control non-treated with IL-6)+/− SEM. Means in non-treated and IL-6 treated groups were compared by student t tests. (D) Hep3B cells were transfected with human <i>HAMP</i> promoter-luciferase construct in combination with either empty vector or human <i>TMPRSS6</i> plasmid. Thirty-two hours after transfection, cells were treated without or with 20 ng/ml of IL-6. After 16 hours of IL-6 treatment, luciferase activity was determined. The assay was performed in duplicate for a total of 6 independent experiments. Values shown are means of luciferase activity divided by a calibrator quantity (the mean value of luminescence for the mock non-treated with IL-6)+/− SEM. Means in non-treated and IL-6 treated groups were compared by student t tests. P values p<0.05 were considered statistically significant. Significances are: *p<0.05.