Salmonella mediates HFE gene transfer in HC macrophages.
Expression of different genes in HC macrophages as detected by RT-PCR at 24 hours (lanes 1 and 4), 30 hours (lanes 2 and 5), and 48 hours (lanes 3 and 6) after Salmonella infection. Lanes 1 to 3, macrophages transfected with βgal; lanes 4 to 6, macrophages transfected with HFE; lane 7, noninfected HC macrophages. The same cDNA preparations were divided and amplified in duplicate test tubes with primers for either HFE and βgal (A) or HFE and HFE/c-myc (B), using β2M primers as an internal control for each reaction. The presence of the transgene is demonstrated by the detection of HFE/c-myc RNA. (C) Western blot of proteins from infected cells. Proteins (30 μg) from cell homogenates were separated by SDS-PAGE, blotted, and analyzed by Western blot using the anti-HFE polyclonal antibody. A significant increase of HFE expression was detected in cells infected with Salmonella carrying the pCMV-HFE plasmid (lane 2) compared with those infected with the pCMV-βgal plasmid (lane 1).
