Fig. 5. Two T-binding motifs and autoregulation of As-T2. (A-D) Expression of lacZ in the 110-cell stage embryos when p(-230)As-T2/lacZ was injected into fertilized eggs without (A) or with As-T2 mRNA (B,C) or HrBra mRNA (D). (B) As-T2 mRNA was tagged with a sequence encoding GFP, and the expression of GFP in embryos showed the proper translation of As-T2 mRNA. (E-H) Expression of lacZ in the 110-cell stage embryos when p(-351)As-T2/lacZ was injected into fertilized eggs alone (E) or with As-T2 mRNA (F,G) or HrBra mRNA (H). (F) GFP expression confirming proper translation of injected As-T2 mRNA. The red box indicates the putative minimal promoter of As-T2; and Td indicates the distal and Tp the proximal T-binding motif. Injected eggs were allowed to develop to the 110-cell stage, and then cleavage was arrested for about 12 hours before detection of the reporter gene expression. Scale bar: 100 µm.