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Fig. S5 Transgene expression is temporally and spatially regulated like the endogenous gene and is dependent on Spt and Ntl function. (A) Embryos were injected with the tbx6 transgene with or without spt and ntl MOs and the percentage of embryos expressing mCherry at the margin was recorded at various stages of development. (A) The tbx6 transgene is initially activated at shield stage, as has been reported for endogenous tbx6 (Hug et al., 1997) (unbroken line). Injecting spt and ntl MOs eliminated mCherry expression (broken line). Similar experiments examining transgene expression in dld:mCherry stable lines are shown in Fig. 9. (B-E) Double in situ hybridizations for mCherry (in red) and tbx6 (B,C) and deltaD (D,E) (in blue) were performed at 75% epiboly. C and E are higher magnification pictures of the embryos in B and D. In all cases, the majority of mCherry expression occurred in regions that also expressed the endogenous gene, or where the endogenous gene was recently expressed.