Figure 2.

Functionally assessed TBX4 (T-BOX transcription factor 4) variants by luciferase assay inducing gain or loss of function. (A) Schematic representation of the in vitro dual-luciferase reporter assay. We cotransfected three different plasmids: Firefly luciferase with x3 T-BOX motifs as the promoter, Renilla luciferase, and TBX4 overexpression plasmid (wild type/mutated). (B) Variants inducing gain or loss of function grouped by primary phenotype; lung disease (perinatal-, childhood-, and adult-onset) or small patella syndrome (SPS). The y-axis represents the relative luciferase units; the dashed line marks the level of the wild type. Data are shown as box plots representing median ± quartiles. Dots represent biological replicates with corresponding batches in different shapes. CMV = Cytomegalovirus promoter; Flag = Flag tag; Myc = partial cMyc tag.