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Fig. 9. Surface localization of TfR. Bloodstream 221 cells were cultured 48 hours in HMI9 media containing 10% fetal bovine serum (BS, panels A,D,G) or 10% canine serum (CS, panels B,C,E,F,H,I). (A-C) Fixed cells were stained with anti-TfR antibody and colloidal gold-conjugated secondary antibodies prior to silver enhancement and preparation for electron microscopy. Note that this fixation technique does not allow detection of TfR in the lumen of the flagellar pocket (FP). (D-I) Live cells were incubated with Tf:gold (5 nm) conjugate at 8°C and prepared for electron microscopy as described in Materials and Methods. 1 mg/ml bovine holotransferrin was included in F,I as a competition control for specific binding. Arrowheads indicate endosomal elements containing Tf:gold. Size scales are indicated in µm. Selected regions of panels D, E and F are magnified 2x in panels D', E' and F', respectively.
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