Fig. 5

CCDC25 affects the hippo pathway by promoting YAP phosphorylation through LATS1. A Knockdown efficiencies of LAST1 in 786-0 cells was detected by Western blot analysis. B Knockdown efficiencies of LAST2 in 786-0 cells was detected by Western blot analysis. C YAP phosphorylation was detected by Western blot analysis, using ACTIN as reference. Quantification of protein expression revealed that knocking down LATS1 inhibited YAP phosphorylation, while the effect of LATS2 was relatively low. D Representative immunoblots of biomarkers of the Hippo pathway, using ACTIN as reference. Quantification of protein expression revealed that knockdown of LATS1 significant attenuated the increase of YAP phosphorylation and reduce of ITGA3 and CCND1 expression caused by CCDC25 overexpression. E 293 T cell lysates were prepared by weak RIPA lysis and co-immunoprecipitated by YAP1-Flag with IgG as negative control. The result showed that YAP interacts with CCDC25 and LATS1 and overexpression of CCDC25 promotes the binding of LATS1 to YAP.