Figure 4
From: Activin A induces growth arrest through a SMAD- dependent pathway in hepatic progenitor cells
Knockdown of Smad4 blocks the anti-proliferative effect of activin A in LE6 cells. (A) SMAD4 expression was determined by western-blot in LE6 cells after Smad4 knockdown (sh1, 2, 3, 4), and compared with control cells (wt) and vehicle cells (V). (B and C) Control cells, vehicle and Smad4 knockdown LE6 cells (LE6-Smad4 KD) were treated with or without 200Â ng/ml activin A. Phosphorylated SMAD2 was detected by western-blot (B) and SMAD2/3/4 complex formation was detected by co-immuno precipitation (C). (D) shSmad4-LE6 cells were treated with or without 200Â ng/ml activin A and their proliferation assessed by CCK-8 and BrdU incorporation assay. c: control, v: vehicle, sh3: LE6-smad4KD. (E) Indicated cells were treated with or without 200Â ng/ml activin A, phosphorylated Rb, cyclinD1, cyclinE , p21WAF1/Cip1and p15INK4B were analyzed by western blotting.