Fig. 3

Western blot analysis of RIPPLY3 expression and functional analysis of the RIPPLY3 variants in the inhibition of TBX1 transcriptional activity in vitro. a Variant RIPPLY3 protein showed no significant change in dosage; b, c Wild-type TBX1 transactivated the wnt5a-Luc reporter and FGF10-Luc reporter compared with the empty expression vector, and wild-type RIPPLY3 showed inhibition of TBX1 transcriptional activity. Compared with wild-type RIPPLY3, three RIPPLY3 variants (p.P30L, p.T52S and p.V179D) showed impaired inhibition of TBX1 transcriptional activity in both homozygote and heterozygote. The results are expressed as relative luciferase activity and the presented values are the mean ± standard deviation of three independent experiments carried out in duplicates. *p < 0.05, **p < 0.01, when compared with wild type RIPPLY3