FIG 2.

Intranuclear confocal microscopy analysis of ASP expression in unstimulated U1C8 cells. Cells were fixed and permeabilized with a FoxP3 staining kit to allow detection of nuclear proteins. Then, the cells were stained with Alexa Fluor 647-labeled anti-ASP 324.6 (red), FITC-labeled anti-LBR (lamin B receptor) (green), and DAPI (blue). Finally, the cells were washed, fixed, and analyzed by confocal microscopy as described in Materials and Methods. The cells display a nonuniform, polarized nuclear distribution of ASP. Bars, 5 μm. Panels B, C, E, and F are three-dimensional representations reconstructed from z-stack images 0.7 μm thick. The results shown are representative of at least three independent experiments.