Figure 5
Figure 5. Scl, Fli1, and GATA2 are downstream targets of Etv2. (A) Alignment of candidate binding sequences containing the Ets binding motif. Putative Ets- or Fox;Ets-binding sequences from human and mouse genomes of Scl, Fli1, and GATA2 were aligned. (B) EMSA showing binding of in vitro–translated Etv2 to oligonucleotide probes containing the Ets-binding sites shown in Figure 6A for Scl, Fli1, and GATA2. Binding was competed by excess wild-type but not by Ets motif–mutated probes. (C) ChIP analysis demonstrating the binding of Etv2 to target sequences. F2 ECs were transfected with HA-tagged Etv2 expression vector and subjected to ChIP analysis. Primer set for each analysis was for Scl;Scl-1_F2/R2, Fli1;Fli1F11/R11, and GATA2;GATA2-F3/R3. (D) Expressions of Scl, GATA2, and Fli1 at E7.5 and E8.5 in mouse embryos. Expressions of Scl, GATA2, and Fli1 were examined by in situ hybridization. These 3 genes are expressed in the extra-embryonic mesoderm of E7.5 (top) and along the dorsal aorta of E8.5 embryos (middle and bottom). Scale bar indicates 200 μm.

Scl, Fli1, and GATA2 are downstream targets of Etv2. (A) Alignment of candidate binding sequences containing the Ets binding motif. Putative Ets- or Fox;Ets-binding sequences from human and mouse genomes of Scl, Fli1, and GATA2 were aligned. (B) EMSA showing binding of in vitro–translated Etv2 to oligonucleotide probes containing the Ets-binding sites shown in Figure 6A for Scl, Fli1, and GATA2. Binding was competed by excess wild-type but not by Ets motif–mutated probes. (C) ChIP analysis demonstrating the binding of Etv2 to target sequences. F2 ECs were transfected with HA-tagged Etv2 expression vector and subjected to ChIP analysis. Primer set for each analysis was for Scl;Scl-1_F2/R2, Fli1;Fli1F11/R11, and GATA2;GATA2-F3/R3. (D) Expressions of Scl, GATA2, and Fli1 at E7.5 and E8.5 in mouse embryos. Expressions of Scl, GATA2, and Fli1 were examined by in situ hybridization. These 3 genes are expressed in the extra-embryonic mesoderm of E7.5 (top) and along the dorsal aorta of E8.5 embryos (middle and bottom). Scale bar indicates 200 μm.

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