Supporting data for “The role of TGF- β signaling in retinal angiogenesis and Familial exudative vitreoretinopathy (FEVR)”

In this study, we collected thirteen FEVR patients and conducted whole-exome sequencing, aiming to identify the novel disease genes and underlying pathogenic mechanisms for FEVR patients. After the integrative analysis, we suggested that TGFBR2 could be a candidate disease gene. We knocked down the TGFBR2 expression in the human retinal endothelial cells and then characterized the cell morphology and proliferation changes, investigated the migration capacity through wound healing assay and transwell assay, and observed the in vitro angiogenesis process by tube formation assay, EdU assay, and immunofluorescence staining. We observed impaired proliferation, migration, and tube formation. Mutant TGFBR2 receptors were overexpressed in the HEK293T cells, followed by the quantification of expression and cell localization. These images were taken from these various experiments in human retinal endothelial cells and HEK293T cells. Additionally, the effect of variants on TGFBR2 expression was quantified by the western blot. Cell lysis was also extracted from human retinal endothelial cells and HEK293T cells after various treatments, including knocking down the expression of TGFBR2, ALK1, SMAD2, and p65 or adding proteasome or lysosomal inhibitors. Then, the targeting protein expressions were quantified.