Fig. 6. Recycling of VSG. Wild-type, mca2/3mca5 and triple RNAi lines, with or without induction with tetracycline for 8 hours. Parasites were biotinylated at 4°C prior to incubation at 37°C for 5 minutes in HMI-9 medium. Remaining surface label was then cleaved by incubation with 50 mM glutathione at 4°C. Cells were prepared for fluorescence microscopy both immediately after the glutathione incubation and following a 5- or 10-minute chase period in HMI-9 medium at 37°C. Parasites were stained with Texas-Red-conjugated streptavidin (red). DNA was visualised with DAPI (blue). Insets, DIC images of the parasite. Bars, 10 µm.