Journal: The Journal of Cell Biology

Article Title: Glypican and Biglycan in the Nuclei of Neurons and Glioma Cells: Presence of Functional Nuclear Localization Signals and Dynamic Changes in Glypican During the Cell Cycle

doi:

Figure Lengend Snippet: The location of potential nuclear localization signals in biglycan ( A ) and glypican ( B ) is indicated by the shaded regions, that were used for the construction of β-galactosidase fusion proteins. The clusters of basic amino acids are underlined. In one glypican mutant, the basic cluster was mutated to NSSSAN, and the sequence indicated by italics was deleted in a second mutant. The attachment sites for the phosphatidylinositol anchor and the COOH-terminal heparan sulfate chain of glypican are indicated by an open triangle and an arrow, respectively. In two fusion proteins, amino acids 499–558 of glypican were replaced by the human IgG Fc sequence (shown as hatched regions in C and D ), and in a deletion mutant, the potential nuclear localization signal used for the β-galactosidase fusion protein was removed from the glypican–Fc fusion protein ( D ).

Article Snippet: Monoclonal and rabbit polyclonal antibodies to Escherichia coli β-galactosidase were obtained from Promega (Madison, WI) and 5 Prime—3 Prime, Inc. (Boulder, CO), respectively, and were used for immunocytochemistry at a dilution of 1:500. mAbs to the HSV and human Fc tags were obtained from Novagen (Madison, WI) and Jackson ImmunoResearch Laboratories (West Grove, PA), respectively.

Techniques: Mutagenesis, Sequencing

Journal: The Journal of Cell Biology

Article Title: Glypican and Biglycan in the Nuclei of Neurons and Glioma Cells: Presence of Functional Nuclear Localization Signals and Dynamic Changes in Glypican During the Cell Cycle

doi:

Figure Lengend Snippet: Localization of β-galactosidase immunoreactivity in transfected 293 cells. 293 cells were transfected with β-galactosidase alone ( A and B ) or β-galactosidase fusion proteins containing either the putative nuclear localization signals of glypican ( C and D ) or biglycan ( E and F ). In a third fusion protein, the basic cluster of the glypican fragment was mutated to nonbasic amino acids ( G and H ), and in a fourth fusion protein ( I and J ), there was a further deletion of the amino acids shown in italics in Fig. B . A, C, E, G, and I show propidium iodide staining; B, D, F, H, and J show β-galactosidase immunoreactivity. Bars, 10 μm.

Article Snippet: Monoclonal and rabbit polyclonal antibodies to Escherichia coli β-galactosidase were obtained from Promega (Madison, WI) and 5 Prime—3 Prime, Inc. (Boulder, CO), respectively, and were used for immunocytochemistry at a dilution of 1:500. mAbs to the HSV and human Fc tags were obtained from Novagen (Madison, WI) and Jackson ImmunoResearch Laboratories (West Grove, PA), respectively.

Techniques: Transfection, Staining