Research Paper Volume 12, Issue 20 pp 20047—20068

Figure 5. CUEDC1 interacts with Smurf2 and inhibits TβRI/Smad signaling pathway and EMT in a Smurf2-dependent manner. (A) Interaction between CUEDC1 and Smurf2 from Pathway Commons (http://www.pathwaycommons.org). (B) Immunoblotting analysis of lysates after immunoprecipitation from H460 cells transfected with Flag-CUEDC1. IgG was used as a negative control. (C) Control and CUEDC1-shRNA cells were treated with or without 10 μM MG132 for 6 h. Cell lysates were immunoblotted with the indicated antibodies. (D) The stability of the Smurf2 protein was measured by immunoblots in NSCLC cells treated with or without CHX. For normalization, β-actin expression was used as a control. (E, F) After Smurf2 overexpression (E) or knockdown (F), EMT markers and components of the TβR I/Smad signaling pathway were detected by western blotting.