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Fig. 6. Tbx5 and Tbx20 are localized to the nucleus and can
activate transcription on the Nppa/ANF promoter. (A)
Schematic depicting the amino acid positions of the T-box domains of
Tbx5 and Tbx20. (B) Schematic of Rat
Nppa/ANF-luciferase reporter construct showing
T-box-binding site consensus sequences and their relative position within the
promoter relative to translation start site. (C-H) Transfected cells were
stained with anti-V5 (C,F; Cy2, green) for TBX5-V5 and TBX20-V5, and with
anti-phosphotyrosine (D,G; Cy3, red) to visualize cytoplasmic compartment.
Overlaid image of Cy2 and Cy3 staining (E,H). (I) Rat
ANF-luciferase co-transfected with a constant amount of
Tbx5 (100 ng) and increasing amounts of Tbx20 (25, 50, 100
and 500 ng) in 293T cells, and the level of transcriptional activation is
expressed as relative luciferase units based on average of three independent
experiments performed in triplicate.
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