Fig. 4

Co-immunoprecipitation assays to analysis the effect of RIPPLY3 variants on RIPPLY3-TBX1 interactions. a The whole cell extracts from the HEK293T cells overexpressing either wild-type or variant RIPPLY3 and TBX1 were immunoprecipitated using the anti-TBX1 antibody; b the grayscale value of the Co-IP protein bands was calculated using Image J software, the results are expressed as the ratio of the grayscale value of RIPPLY3 protein band divided by the TBX1, and the wild-type ratio is standardized to 1.0. Co-IP studies showed that the p.T52S significantly disrupted the physical interaction between RIPPLY3 and TBX1, whereas the p.P30L, p.D113N and p.V179D had no effect on the interaction between RIPPLY3 and TBX1 compared with the wild-type. *p < 0.05, when compared with wild type RIPPLY3