Figure 2.

GATA2 mutations impair basal preproTRH transcriptional activity. (A) Schematic representation of rTRH(547)-CAT. GATA-RE; GATA responsive element; AP-1, AP-1-binding site; GRE, glucocorticoid responsive element; GRE1/2, glucocorticoid responsive element half-site; STAT, signal transducer and activator of transcription; CRE, cyclic AMP response element; GCA, GCA site; KEM1, KEM1 site; TSS, transcription start site; CAT, chloramphenicol acetyltransferase. (B) and (C) rTRH(547)-CAT (1.0 μg) was transfected into CV1 cells along with expression plasmids for various GATA2 mutants (0.1 μg), TRβ2 (0.1 μg), and pCMV-β-galactosidase (0.2 μg). After 24 h of culture, the cells were treated with vehicle or 10 nM T₃ for an additional 24 h. CAT activity was normalized with β-galactosidase activity. CAT activity for pCMV-CAT (5 ng/well) was taken as 100%. Data are expressed as the mean ± S.E. of at least three independent experiments. * p < 0.05; *** p < 0.001, compared with wild-type (WT). §, p < 0.05 for the indicated comparison.