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Fig. 1. Characterization of CFP-vimentin and YFP-ß3 integrin expression in TrHBMEC transfectants. (A) Extracts of transformed human bone marrow endothelial cells (TrHBMECs) co-transfected with expression vectors encoding CFP-vimentin and YFP-ß3 integrin were processed for western immunoblot analysis using antibodies against GFP (lane 1), ß3 integrin (lane 2) or vimentin (lane 3). Molecular masses are indicated on the left. The reactive species are indicated on the right. (C-I) Images of transfected TrHBMECs. Cells expressing CFP-vimentin (B,F) and YFP-ß3 integrin (C,G) were fixed and then processed for immunofluorescence microscopy using an anti-ß3 integrin subunit antiserum (D) or anti-vimentin antibodies (H). E and I are the merged images of B-D and F-H, respectively. YFP-ß3 integrin (C) colocalizes precisely with the endogenous {alpha}vß3 integrin complexes in FC (D) (purple in the merged image in E). CFP-vimentin (B) colocalizes precisely with the endogenous vimentin intermediate filaments (H) (sky blue in the merged image in I). Scale bar: 10 µm. Vim, vimentin.