Antibody staining was done on ice. Cells were washed once with PBS/1% FCS and afterwards incubated for 30 minutes in a total volume of 100 μl with the respective recommended amount of antibody. Following antibodies were used in our study: anti-CD71 (BDPharmingen; M-A712; APC), anti-CD28 (BDPharmingen; L293; PE), anti-HFE (Abnova; polyclonal), anti-CD3 (Caltag; UCHT1; APC), anti-CD4 antidody (Caltag; RPA-T4; APC), anti-HLA-ABC (Dako; W6/32; PE). Post staining cells were washed twice and fixed with 2% PFA/PBS. At least 2.000 infected cells were measured with a FACS Canto II (BDBioscience). Fold modulation of cell surface receptor modulation was calculated as before [14 (link),51 (link)]. Flow cytometric measurement of FRET and the according gating strategy were performed as already described [24 (link)]. Cellular iron content was measured by the use of the fluorescent dye CA-AM which is quenched by free cellular iron [4 (link)].
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