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Fig. 1. The effect of the gypsy sequence on the nuclear localization of neighboring chromosomal DNA. (A-C) Confocal images of Drosophila imaginal disc nuclei. Disc tissue was visualized with FISH using Cy3-labeled probes (red) and an antibody against the nuclear lamin (green). The main view panel and the side view bars are separated by white lines. The positions of focal planes are indicated by thin yellow lines. Nuclei from non-suHw-LacZ (NS) transgenic (A) or non-transgenic (B) larvae were hybridized with Cy3-labeled LacZ probe. FISH foci were visible within nuclei from transgenic larva. In the majority of the nuclei from the non-transgenic larvae, no stained foci were observed above background. One or sometimes two fluorescence foci per nucleus were detected at various focal planes in virtually all nuclei, depending on transgene copy number and the status of chromosomal pairing between the homolog transgenes. A small number of Cy3-labeled foci appear outside of the lamin ring due to weak or incomplete lamin staining of the nearby nuclei. Arrowheads in A and C indicate Cy3-labeled foci that appear in the center of the nucleus in the top view but are more peripheral in the side view. (C) Nuclei from a gypsy line were hybridized with a Cy3-labeled probe for an 8 kb genomic region in the cut locus near the retrotransposon (gypsy-Cy3). The gypsy foci (red) are frequently present near the nuclear periphery (green). (D) Histograms of 2D foci-distribution of individual NS transgenes (colored lines) and of 3D foci distribution of pooled NS transgenes (black line). Relative frequency of the sample is plotted against the FISH-lamin distance (% radius) of the sample. (E) Histograms of 2D foci distribution of individual gypsy lines (colored lines) and of 3D foci distribution of all gypsy lines (black line). (F) Summary of foci distribution in a quartile diagram. The relative FISH-lamin distance of each sample group (colored bars) is compared with a model of randomly distributed samples (gray bar) (see Materials and Methods for details). (G) Foci histograms along the nuclear radius of selected sample groups (colored lines) and the probability density function along the nuclear radius for the random distribution model (gray dashed line) (see Materials and Methods).
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