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Fig. 2. Comparison of Cre recombinase activity in the developing forelimbs of Prx1Cre and Prx1CreERT2 transgenic embryos using the Rosa26RlacZ reporter line. CreERT2 was activated by gavaging the pregnant females at E8.5. Activation of the reporter by Cre recombination is detected by lacZ staining (blue). In Prx1Cre embryos, Cre activity was detected throughout the FL-forming region at the 14-somite stage (A), and was induced in all cells of the FL of 19- and 21-somite stage embryos (B,C). In Prx1CreERT2 embryos, Cre activity was also first detected at the 14-somite stage, but only in a small number of cells (D,G). Low level activity was still apparent in 16- to 17-somite stage embryos (E,H), and it was not until the 21-somite stage that Cre activity was detected throughout the limb bud (F). By E10.5, the Cre reporter was uniformly activated throughout the limb bud (I-K). When the Prx1CreERT2 transgenic was activated in a background of the Tbx5 conditional allele (Tbx5^lox/lox) and Cre reporter, a bud formed despite uniform Cre activity throughout the limb (L,M). PCR was used to quantitate the levels of Tbx5 ex3 loss through recombination following TM gavage at E8.5. Recombination of 87%-97% was observed in two independent E10.5 Prx1CreERT2; Tbx5^lox/lox FLs (Mut1 and Mut2) (N). The graph shows the mean values and s.d. obtained from three replicates, normalised to a control; the mean value obtained from the wild-type limb is shown as 100%.

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