Fig. 3

Interaction between Tbx5 and trisomic gene, Adamts1. Quantitative PCR of Adamts1 mRNA levels in pooled E11.5 hearts (a). Adamts1 expression levels are significantly increased in euploid Tbx5 ^{+/ -}mice (1.95x, p = 0.04). No other changes were significant compared to the wild type expression. Error bars are standard deviation from the mean. TBX5 binding upstream of the Adamts1 locus alters transcription (b). Cells transfected with the Adamts1-luc construct showed increased luciferase when compared to the pGL3 construct (p = 1.28 × 10⁻⁴). Cells transfected with the Adamts1-luc construct and Tbx5 siRNA showed significantly lower luciferase levels when compared to those transfected with the Adamts1-luc construct alone (p = 3.35 × 10⁻⁴, compared to Adamts1-luc). Luciferase levels dropped significantly when the TBE was mutated to inhibit TBX5 binding (p = 1.51 × 10⁻⁴). Addition of Tbx5 siRNA did not affect transcription when the TBE was mutated (p = 0.9) Error bars are standard deviation. The region inserted into the basic pGL3 vector is 121 bp upstream of the Adamts1 gene, contains a putative TBE and associated with TBX5 in a ChIP study. Tbx5 over expressing cells were harvested 48 h after transfection. The FGF10-luciferase construct was used as a positive control. Graph represents one experiment representative of all replicates