Figure 1 uPA variants translocate to the... | American Society of Hematology

Figure 1

$Figure 1. uPA variants translocate to the nucleus. HeLa cells (A) or BJ cells (B) were incubated for 30 minutes with 10 nM WT-scuPA or ΔGFD-scuPA, respectively, in complete medium at 37°C, washed, and fixed in cold MeOH for 15 minutes. To visualize uPA, the fixed cells were incubated with α-uPA pAb and Alexa488-conjugated α-rabbit pAb. Nuclei were stained with DAPI. Images were taken using a Zeiss LSM 510 laser scanning confocal microscope with the z interval set at 0.3 μm. (i-iii) Confocal images taken through a single 0.3-μm optical slice. (i) Staining with α-uPA Ab is shown in green. (ii) Counterstain of the nucleus with DAPI is shown as pseudocolor in red. (iii) Merged images within one focal plane. Scale bar represents 10 μm. (iv) Sequences of merged images taken in various z sections at 0.3-μm intervals were subjected to 3-dimensional (3D) reconstitution using Volocity 4.1.0 software (Improvision, Lexington, MA). Virtual 3D image of the cell was sectioned along the XZ plane through the nucleus. The results reveal that a substantial fraction of scuPA and/or ΔGFD-scuPA is found within the nucleus.$

uPA variants translocate to the nucleus. HeLa cells (A) or BJ cells (B) were incubated for 30 minutes with 10 nM WT-scuPA or ΔGFD-scuPA, respectively, in complete medium at 37°C, washed, and fixed in cold MeOH for 15 minutes. To visualize uPA, the fixed cells were incubated with α-uPA pAb and Alexa488-conjugated α-rabbit pAb. Nuclei were stained with DAPI. Images were taken using a Zeiss LSM 510 laser scanning confocal microscope with the z interval set at 0.3 μm. (i-iii) Confocal images taken through a single 0.3-μm optical slice. (i) Staining with α-uPA Ab is shown in green. (ii) Counterstain of the nucleus with DAPI is shown as pseudocolor in red. (iii) Merged images within one focal plane. Scale bar represents 10 μm. (iv) Sequences of merged images taken in various z sections at 0.3-μm intervals were subjected to 3-dimensional (3D) reconstitution using Volocity 4.1.0 software (Improvision, Lexington, MA). Virtual 3D image of the cell was sectioned along the XZ plane through the nucleus. The results reveal that a substantial fraction of scuPA and/or ΔGFD-scuPA is found within the nucleus.

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