Research Paper Volume 11, Issue 21 pp 9719—9737

Figure 7. Tan IIA destabilized YAP through regulating SMAD7 in liver cancer. (A, B) Colocalization of SMAD7 and YAP in Bel-7404 cells and xenograft tissues analyzing by immunofluorescence assay. Scale bar: 50 μm. (C) Western blot assay analysis of SAMD7 and YAP protein expression in Bel-7404 and SMMC-7721 cells transfected with different dose SMAD7-Flag. (D) YAP and SMAD7 protein expression level were analyzed by western blot assay treated with or without MG132 in different groups. (E) P-YAP, YAP and βTrcp1 protein expression level were measured by western blot assay with or without SMAD7-Flag overexpression in Bel-7404 and SMMC-7721 cells. (F) Relative YAP/P-YAP ratio was measured by gray value. ***p<0.001 vs control. (G, H) Bel-7404 cells were stably transfected with or without SMAD7-Flag, with or without Tan IIA treated, and protein synthesis was blocked by treatment of CHX for the different time points. Relative SMAD7 and YAP protein levels were detected by western blot assay.