Fig. 2

DYRK1 bound with and phosphorylated endophilin. A The workflow for screening DYRK1 substrates. Phosphoproteomics sequencing of DYRK1 inhibitor-treated samples identified the downregulated phosphorylation proteins. The potentially interacting proteins of DYRK1 were screened by a Y2H screening assay. Endophilin is a member of overlapped proteins by two technique approaches. B Subcellular localization of endophilin. It was shown by immunofluorescence that the endophilin was localized at the center of the anterior and posterior membrane regions where the lumen will appear (white arrowheads) at 18 hpf (the initiation of lumen formation). At 22 hpf (lumen expansion), endophilin was mainly enriched in the apical membrane (white arrowheads). The region of white box was magnified for observing localization. C Interaction assays between DYRK1 and endophilin by Y2H. Both kinase-active and inactive forms of DYRK1 interacted with endophilin. D Interaction of DYRK1 with endophilin was confirmed by Co-IP assay in the HEK293T cell line. Scale bar 10 μm