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. 2015 Jun 11;5:10394. doi: 10.1038/srep10394

Figure 1.

Figure 1

Manipulation of the size of VSG by expression of fusion proteins and chemical labelling. a) The VSG coat of chemically fixed bloodstream form trypanosomes is recognized by VSG-specific primary antibodies and green fluorescent secondary antibodies. b) Fusion of the green fluorescent protein (GFP) to the N-terminus of VSG M1.2 produces an enlarged VSG chimera (VSG-GFP; right). The transgenic trypanosomes reveal a strong green fluorescence, which, however, is restricted to the biosynthetic and endosomal compartments. No cell surface fluorescence is detected (left). c) Fusion of the C-terminal domain of VSG M1.2 to GFP (tVSG-GFP). From structural homology modelling (right) the resulting chimeric protein is expected to be significantly smaller than the native VSG. The fusion protein is expressed, however, it is also retained intracellularly (left). d) Live cell labelling of biotinylated trypanosomes with Alexa 488-conjugated streptavidin (left). The streptavidin molecule enlarges the VSG by 5 × 5.5 nm (right) e) Live cell labelling of trypanosomes with sulfo-NHS-Atto488. The small, membrane-impermeable fluorophore evenly tags the VSG coat of living trypanosomes. Scale bars: 3 μm.