Fig. 5: JOSD1 is associated with YAP and regulates YAP protein stability.

A Immunofluorescence staining was used to analyze the intracellular localization of JOSD1 and YAP in HCT116 cells grown in standard medium. The findings show the localization of JOSD1 (green) and YAP (red), while nuclei are stained blue with DAPI. B In HCT116 cells, JOSD1 and YAP were primarily located in the nucleus. C In HCT116 cells, the Co-IP assay showed that endogenous JOSD1 and YAP are associated. An antibody was used for Co-IP. D Knockdown of JOSD1 does not result in further degradation of YAP in the presence of the proteasome inhibitor MG132. E When MG132, a proteasome inhibitor, was present, JOSD1’s stabilizing effect on YAP did not cause an additional increase in the YAP protein level. F, G In HEK293T cells, overexpressing JOSD1 extended the half-life of YAP, whereas overexpressing JOSD1^C36A did not. H, I Depletion of JOSD1 reduced the half-life of YAP protein in HCT116 cells. The halftime of YAP protein in (I) was quantitatively analyzed. J, K Schematic diagrams showing the wild-type and truncated YAP and JOSD1 constructs. L, M Immunoblots display the JOSD1 interaction with WT or truncated YAP through immunoprecipitation with JOSD1 (anti-Flag), and YAP interaction with WT or truncated JOSD1 via immunoprecipitation with YAP (anti-Myc). All data are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 by one-way ANOVA.