Cell surface expression of HFE in transfected macrophages.

Human macrophages from a patient with HC were infected in vitro withSalmonella strain SL7207 carrying either the pCMV-βgal (middle panels) or the pCMV-HFE (lower panels) plasmid. Results from a representative experiment in which cells were analyzed for gene expression by FACS 48 hours after infection using monoclonal antibodies against major histocompatibility class I, HFE, CD71, and βgal followed by a PE-conjugated anti-murine IgG polyclonal antibody. Appropriate isotype-matched monoclonal antibody controls have been used for instrument settings (empty histograms). Detection of βgal expression was performed after saponin-mediated permeabilization of monocytes. Percentage of positive cells and mean values of fluorescence (in brackets) are reported for each single histogram. Basal expression level for each marker in untreated HC–monocytes is also shown (upper panel).