Fig. 6

The Zebrafish Enhancer Detection (ZED) vector effectively detects enhancer activity. For each construct, the number of injected embryos was higher than 100. A: Green fluorescent protein (GFP) expression in 24 hours postfertilization (hpf) mosaic F0 embryos injected with the ZED vector harboring the E187, E200, E261, E298, Z176 and Z48 enhancers and F1 embryos derived from the Z176 and Z48 constructs. Insets show GFP and red fluorescent protein (RFP) expression in these two last F1 stable lines at 48 hpf. Note the reduced background in these injected embryos as compared to those injected with the noninsulated pgata2 constructs shown in Supp. Fig. S1. For the F1 lines, we obtained more than two independent insertions that showed the same pattern. B: Graphic representation of the percentage of embryos showing GFP expression in the expected territories. The reduced background observed with the ZED constructs did not compromise the enhancer detection potential of this vector as the GFP activity was detected in 30-50% of the injected embryos (n > 100). These percentages are similar to that observed in embryos injected with the noninsulated pgata2 constructs containing the same enhancers (Supp. Fig. S1).