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. 2015 Sep 22;290(47):28107–28119. doi: 10.1074/jbc.M115.662197

FIGURE 3.

FIGURE 3.

Autoactivation of Etv2 gene expression. A, endogenous Etv2 expression in iHA-Etv2 ES differentiation on day 4 upon doxycycline (Dox) induction for 6 h is increased and inhibited after 24-h treatment. *, p < 0.05. B, schematic of the ChIP strategy. Primer pair F1 and R1 flanks the Ets#2 and Ets#3 motifs, and F2 and R2 flanks the Ets#4 motif. C, ChIP analysis of the interaction between Etv2 and the Etv2 promoter using iHA-Etv2 EBs on day 4. Etv2 preferentially binds to CRII and less so to CRI. *, p < 0.05. D, Etv2 transactivates the Etv2 minireporter ∼8-fold in 293T cells. Mutation of the Ets#1 or Ets#4 motif has no effect on transactivation, whereas mutation of the Ets#2 or Ets#3 motif results in loss of transactivation. Mutation of all four Ets motifs has the same effect as that of the Ets#2 or Ets#3 motif mutants. *, p < 0.05. E, both Etv2 and Gata2 transactivate the Etv2 minireporter in 293T cells ∼6-or 2-fold, respectively. Together, Etv2 and Gata2 transactivate the Etv2 minireporter to ∼10-fold. *, p < 0.05.