Fig. 6

Co-immunoprecipitation of RANK and HER2. a Immunoprecipitation (IP) against HER2 was performed in HEK293 cells transfected with RANK-V5 and HER2, HER2-FLAG, a carboxy-terminal fragment of HER2 (611-CTF) or an amino-terminal fragment of HER2 (742-NTF). IP was performed using anti-FLAG, anti-HA, or control IgG antibodies as indicated. RANK was detected by blotting the immunoprecipitates (IP, left upper panel) or the whole lysates (input, right upper panel) with the V5 antibody. HER2 was detected in IPs (left lower panel) and input (right lower panel) using the 32H2 antibody that detects all forms of HER2. b IP against RANK-V5 was performed in HEK293 cells transfected with RANK-V5 and GFP, HER2-FLAG, a carboxy-terminal fragment of HER2 (611-CTF) or an amino-terminal fragment of HER2 (742-NTF) using the V5 antibody. In the IP and input, HER2 was detected using the 32H2 antibody. c IP against endogenous HER2 was performed in SKBR3 cells using trastuzumab (Herceptin-HCP) or a control IgG. Endogenous RANK and HER2 were detected in IP (RANK immunoprecipitated by HER2 is indicated by an asterisk (*) in the upper panel) and input samples