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. 2005 May;73(5):2690–2697. doi: 10.1128/IAI.73.5.2690-2697.2005

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Copyright © 2005, American Society for Microbiology

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FIG. 5. — LouTat 1 VSG and 117 VSG colocalize to the endoplasmic reticulum, flagellar pocket, and cell surface in LT1:117 VSG gene double expressers. G418-selected trypanosomes were fixed with 1% formaldehyde/0.05% glutaraldehyde and attached to slides by acetone/methanol fixation. Slides were stained with a mouse anti-LouTat 1 VSG monoclonal antibody and a rabbit anti-117 polyclonal serum. Positive cells were detected using Alexa 633-conjugated anti-mouse IgM (red) and Alexa 488-conjugated anti-rabbit IgM (green) and viewed by fluorescent microscopy. Immunostaining: (A) mouse anti-LouTat 1 VSG; (B) rabbit anti-117 VSG; (C) merged image; (D) differential interference contrast with DAPI counterstain. Control stains demonstrated that these antibodies do not cross-react with the heterologous VSG in this assay. Images were pseudocolored and merged using OpenLabs 3.0 software (Improvision, Inc., Lexington, MA).