Fig. 5. Quantitative analysis of the activity of PTS. (A) Representative embryos showing IAB5-lacZ promoter targeting. Red circles indicate the areas measured for relative staining intensity. (B) The graph displays staining intensity as a function of time. We detected almost no staining in the first 7 minutes after adding the substrate NBT/BCIP. This may be due to the time needed for the substrate to diffuse into the embryos. Between 10 and 60 minutes, staining intensity (2^EV-1) was linear with staining time. (C) To compare IAB8 enhancer strength in different P-elements, heterozygous embryos were fixed and stained in parallel for 45 minutes. Approximately 30 embryos were measured. Lanes 1 and 2 show the staining intensities for the IAB8 enhancer on the lacZ promoter when it is located about 100 bp (lane 1, gray), or 5.5 kb (lane 2, green) away from the lacZ promoter. Five strains for each of these transgenes were analyzed. Lane 3 (blue) and 4 (yellow) indicate IAB8-w interaction (W32-w) before and after the removal of PTS and Fab-8 in W32. Lane 5 and 6 shows IAB8-lacZ interaction (W32-Z) in the same strain before and after the removal of PTS and Fab-8. (D) Analysis of enhancer activities before (blue) and after (yellow) the removal of PTS and suHw from W81. Four different transgenic strains (W81-1, W81-2, W81-3 and W81-4) were shown.

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