Fig. 3. Subcellular localization of S100 proteins in endothelial cells (ECV). Several S100 proteins were found in ECV, and different subcellular distributions of the proteins was observed by staining with the appropriate antibodies. Rabbit anti-S100 antibodies were used at a 1:1000 dilution, and a goat Cy3-conjugated anti-rabbit antibody was used for visualization under the 40x oil objective lens in the Zeiss light microscope with a filter of 450 to 490 nm. Nuclear localization was observed in cells stained with S100A13 (A), S100A1 (C), S100A4 (E) and S100A5 (F) antibodies; perinuclear stainings were observed with S100A6 (B) and S100B (G) antibodies. A very faint staining was seen with the S100A3 antibody (D). H shows one example of background staining with pre-absorbed S100A6 antibody, and no obvious staining was observed. The same observation was made with S100A13 and the other pre-absorbed S100 antibodies, indicating the specificity of the antibodies used. Bar, 15 µm.

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