Research Paper Volume 11, Issue 21 pp 9719—9737

Tanshinone IIA mediates SMAD7-YAP interaction to inhibit liver cancer growth by inactivating the transforming growth factor beta signaling pathway

Figure 4. SMAD7 can inhibit liver cancer growth and migration. (A) The bright images of Bel-7404 and SMMC-7721 cells expressing GFP (Control) or SMAD7-Flag. Cells were plated at a density of 5,000 per well and imaged 7 days later (left panel). Scale bar: 100 μm. Cells were then quantified using ImageJ software and the relative cell number are shown as Mean ± SD from three independent tests (right panel). *p<0.05, **p<0.01 vs control. (B) Bel-7404 cells were infected with control or SMAD7-Flag virus as indicated, and cell migration ability was assessed by cell scratch assay. The images are taken at the 0 h, 12 h and 24 h time points after the cultures were wounded by scratching (left panel). Quantification of relative scratch area is measured by ImageJ software, datas are showed as Mean ± SD of three independent experiments (right panel). **p<0.01, ***p<0.001. (C) SMAD7-Flag induced apoptosis marker cleaved caspase substrate expression measured by immunofluorescence assay in Bel-7404 and SMMC-7721 cells. Scale bars: 100 μm. (D) Representative images of control (expressing GFP tag) and SMAD7-Flag (expressing GFP tag) Xenografts mouse were analyzed by small animals in vivo imaging system (left panel) and representative images of tumor as indicated (right panel). (E) Representative HE and IHC images of SMAD7, proliferation marker Ki67 and apoptosis marker Bcl2 staining were taken in control and SMAD7-Flag Xenografts mouse tissue groups at 400Χ magnifications.