(A) The hD2 promoter is synergistically activated by GATA2 and PKA signaling. A CAT-reporter assay was performed as described in Fig 1B in the presence or absence of forskolin (FSK) (10 μM). CAT activity for pCMV-CAT (5.0 ng/well) was taken as 100%. Data are expressed as the mean ± S.E. of at least three independent experiments. *, P<0.05 for the empty vector vs. pcDNA3-mGATA2. (B) TRH-R signaling has modest effect on GATA2-induced transcription of the human D2 promoter. After CV1 cells were transfected with 2.0 μg hD2-CAT along with 0.4 μg pcDNA3-mGATA2 and expression plasmid for TRH-R, cells were incubated with 100 nM TRH for an additional 24 h. *, P<0.05 for the empty vector vs. pcDNA3-mGATA2. N.S., statistically not significant. (C) A Gel-shift assay was performed as described in Fig 3C. Solid arrowhead, GATA2 monomer; arrow, GATA2 dimer; open arrowhead, free probe; SS, super-shift of GATA2 by the anti-GATA2 antibody.