Figure 5

Expression of ferroportin and effect of hepcidin in cultured THP-1 and CEM cells. THP-1 cells (A) or CEM T cells (C) were treated with FAC for 24 hrs and then treated with 100 μM ascorbic acid for 2 hours. The cells were then infected with VSVG-pdseudotyped HIV-1 Luc virus for 48 hrs (A and C, lanes 2 and 3). Lane 3, the cells were treated with 0.5 μM hepcidin for 6 hours. The cells were lysed, using Luclite buffer system and the Luciferase activity was measured using Labsystems Luminoskan and the data normalization to the protein concentration. THP-1 cells (B) or CEM T-cells (D) were treated with FAC for 24 hrs and then with 100 μM ascorbic acid for 2 hours (lane 2). The cells were then treated with 0.5 μM hepcidin for 6 hours (lane 3). The cell lysates were resolved on 10% SDS-PAGE and immunoblotted with anti-MPT1 antibodies or with anti-tubulin antibodies. Lane 1, untreated control cells.