Research Paper Volume 11, Issue 10 pp 3298—3314

Figure 5. Overexpression of GDF10 induces cell cycle arrest and inhibits EMT in BT-20 cells. (A) The expression of cyclin D1, γH2AX, Bax, and active caspase-3 was investigated via western blotting in BT-20 cells transfected with GDF10 for 72 h. β-actin was used as internal control. Relative expression data were quantified by densitometry and normalized to β-actin. **P < 0.01, compared with the NC group. (B) Cell cycle distribution in BT-20 cells transfected with GDF10 for 72 h. *P < 0.05, **P < 0.01, compared with the NC group. (C) Smad7, p-Smad2, Smad2, Smad4, E-cadherin, N-cadherin, and Vimentin expression was assessed by western blotting in BT-20 cells transfected with GDF10 for 72 h. β-actin was used as internal control. Relative protein expression levels were quantified by densitometry and normalized to β-actin. *P < 0.05, **P < 0.01, compared with the NC group. (D) Immunofluorescent assessment of the subcellular distribution of Smad4 in BT-20 cells transfected with GDF10. (E) Smad4 expression was assessed by western blotting in the nucleus and cytoplasma of BT-20 cells transfected with GDF10 for 72 h, respectively. Histone H3 and β-actin were used as internal control respectively. Magnification × 400.