Fig. 1

Identification of YOD1 substrates in the Hippo signaling pathway-related proteins using informatics tools. A A YOD1 interaction network was generated with STRING. The thickness of the line indicates the strength of association. The color of nodes indicates the confirmation of binding between YOD1 and proteins (gray, unpublished; blue, unpublished and related to the Hippo signaling pathway; and green, published and related to the Hippo signaling pathway). USP21 binding with YOD1 is shown in purple. B Myc-YOD1 alone or in combination with Flag-USP21 was transfected into HEK293T cells, IP with an anti-Flag or an anti-Myc antibody, followed by immunoblotting with antibodies against Myc and Flag. C Purified GST or GST-YOD1 was incubated with Flag-USP21-overexpressed HEK293T cell lysates. Purified GST and GST-YOD1 were visualized using Coomassie Brilliant Blue R/G staining solution. D ICC was performed to investigate the localization of respective YOD1 and USP21, and co-localization of YOD1 and USP21 (red, YOD1; green, USP21; and blue, DAPI). Scale bar, 20 μm. E Schematic representation of Flag-USP21 and Myc-YOD1 and their deletion mutants. F Myc-YOD1 and Flag-USP21 or its deletion mutants were co-transfected into HEK293T cells, followed by IP with an anti-Myc antibody and subsequent immunoblotting with Myc and Flag antibodies. G Flag-USP21 and Myc-YOD1 or its deletion mutants were co-transfected into HEK293T cells, followed by IP with an anti-Flag antibody, and subsequent immunoblotting with Myc and Flag antibodies