Figure 1.

NPAT concentrates at a few foci in the nuclei of human cells. (A) Localization of NPAT in the indicated cell lines was examined by indirect immunofluorescence (IF) using an affinity-purified rabbit anti-NPAT antibody (R48). The antirabbit IgG secondary antibody was conjugated with Texas Red (Red). Nuclear DNA (nucleus) was stained with DAPI (blue). (B) Multiple NPAT antibodies recognize the same NPAT foci. WI38 cells were double stained with a mouse monoclonal NPAT antibody (DH3 or DH4) and an affinity-purified rabbit polyclonal NPAT antibody (R48 or R49). The staining from the mouse antibodies (green) and that from the rabbit antibodies (red) are completely overlapping (yellow). (C) Most WI38 and MRC5 cells have two or four NPAT foci (spots). WI38 and MRC5 cells were stained with R48, and the number of NPAT spots in each cell was counted. The mean results from three independent experiments and the standard deviations are shown. More than 200 cells were analyzed in each experiment.