FIG. 5.

FISH analysis of retroviral vector integration. (A) Imaging of nuclei from transduced cells. Cells were infected with the lentiviral vector pSINgfp and cultured for 2 months before FISH analysis. Nuclei were counterstained with propidium iodide. Individual integrated events are revealed by hybridization with a specific Alexa Fluor 488-labeled probe. (B) Peripheral distributions were modeled by random distribution in shells whose thickness was nR/10 (1 < n < 10). (C) Reconstruction of nuclear volume from a 2D image. The sphere (R radius) was fitted by a set of cylinders (each cylinder was defined as the mean radius = r_i, height = h_i, and thickness = ɛ, with ɛ = R/10). In the example shown in the figure, h_i and r_i refer to the third cylinder starting from the center of the sphere. (D) Model of signal distribution in shells of different thicknesses relative to the distance from nuclear border.