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Fig. 3. IFN- induces upregulation of MVP mRNA and protein expression. (A) mRNA contents of Hep3B cells treated with IFN- (250 IU/ml) for 6 hours, 24 hours and 48 hours were analyzed by northern blots in comparison to ß-actin and GAPDH mRNAs as controls. Signals for MVP were normalized to the 28s RNA band and are given as fold induction compared with untreated control cells. (B) MVP expression of human cancer cell lines treated for 24 hours and 48 hours with IFN- (250 IU/ml) were analyzed by western blot. ICAM-1 was used as a positive control for the IFN- response, and ß-actin as a control for equal loading. (C) A representative western blot experiment (out of four experiments performed) as shown in B was quantified using ImageQuant software. Changes in MVP expression by IFN- are given as fold induction compared with control cells at 24 hours. (D) Western blot analysis of protein lysates obtained from cells pretreated for 30 minutes with DMSO (Co) or the indicated concentrations of curcumin or AG490 as indicated followed by incubation without or with IFN- (250 IU/ml) for 24 hours. MVP expression is compared with ICAM-1 as a positive control for the IFN- response and ß-actin to demonstrate equal loading. (E) Western blot analysis was performed using whole cell extracts derived from Hep3B cells transiently transfected with a DN-STAT1 expression or control plasmid. Forty-eight hours after transfection, cells were treated with 250 IU/ml IFN- for 24 hours. All experiments shown in this figure were performed at least twice with comparable results.

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