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Fig. 3. The suHw insulator can function in the nuclear interior. (A-D) Serial confocal sections (0.3 µm thick) of larval imaginal disc cells stained with anti-SUHW (red) and anti-lamin (blue) antibodies. The SUHW protein foci are detected at both the nuclear periphery and the interior region. (E) SUHW protein foci histograms along the nuclear radius. (F-H) A confocal section of an imaginal disc containing the 3S2 transgene, triple stained for the transgene (FISH, red; SUHW, green; lamin, blue). (I-K) Heat shock does not disrupt enhancer-blocking activity of the suHw insulator. Whole-mount-RNA in situ hybridizations were performed with embryos containing 3T2 (I), and 3S2 transgenes (J,K). Transgene constructs are shown below the embryos. Colored bars represent the even skipped stripe enhancers (E2 and E3) and the neutral spacer (T), the black oval represents the 340 bp suHw element (S). Enhancers that directed the lacZ stripe expression are indicated above the embryos. Embryos were incubated at 25°C (D,E) or 37°C (F) for 30 minutes before fixation and staining.





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