Methods & Materials

Generation of Fpn-GFP
A cDNA for mouse ferroportin (Fpn) was cloned. The plasmid was then
digested to obtain the Fpn-GFP construct. A vector was digested and
ligated with Fpn-GFP . The ligation was transfected into E.coli. and
ampicillin-resistant colonies selected. The plasmids were then purified and
sequenced.
Construction of EcR293 Fpn expressing cell line
Cells were transfected with 5 μg DNA using Lipofectamine. After
transfection, the cells were placed into selective medium. 2-3 weeks
later, stably transfected colonies which showed induction of Fpn-GFP in
response to ponasterone were selected.
Transient transfection
A mouse Fpn cDNA was cloned into the sites of a CMV-containing the vector.