Fig. 2

COPS3 interacts with Raf-1 and activates MEK/ERK signaling. a The COPS3 related protein–protein interaction network based on Human Protein Reference Database was constructed using Cytoscape. b 143B or HOS cells lysates were precipitated with COPS3 or Raf-1 antibody followed by immunoblotting probing with the indicated antibodies. c 143B and HOS cells were stably transfected with shCOPS3 or shCtrl lentivirus, COPS3 expression was analyzed by western blotting and Real-time RT-PCR, Raf-1 expression was determined by immunoblotting. d SAOS-2 cells were transfected with COPS3 adenovirus, the COPS3 level was analyzed by western blotting and Real-time RT-PCR, Raf-1 expression was determined by immunoblotting. e 143B and HOS cells stably transfected with shCOPS3 or shCtrl were treated with or without MAPK/ERK inhibitor U0126 (1 μM) for 24 h. Indicated proteins were detected by immunoblotting. f SAOS-2 cells transfected with Ad-COPS3 were exposed to MAPK/ERK inhibitor U0126 (1 μM) for 24 h. Western blot assay was performed with the indicated antibodies. The data are presented as mean ± S.D. from three independent experiments. **P < 0.01 vs. shCtrl group (c) or the Ad-NC group (d)