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. Author manuscript; available in PMC: 2015 Apr 24.
Published in final edited form as: Cell. 2014 Apr 24;157(3):595–610. doi: 10.1016/j.cell.2014.03.027

Figure 1. PTEN exists in a dimeric complex.

Figure 1

(A) Co-IPs from total lysate of PC3 cells. IP of MycPTEN (top panel), and IP of GFPPTEN (bottom panel) with specific tag antibodies revealed reciprocal PTEN-PTEN interaction.

(B) Co-IPs from total lysate of 293T cells, transfected with GFPPTENWT and IP with anti-GFP antibody. Western blot was probed with a PTEN antibody.

(C) NIH3T3 cell lysates were IP with anti-rabbit PTEN antibody; native elution and Western blots show monomer and dimer of the protein using mouse anti-PTEN antibody.

(D) In bacteria, GST-PTEN specifically pulls down His-PTEN.

(E) Renilla and GFP emissions are detected at the indicated wavelengths. Administration of Coelenterazine induces Renilla excitation which generates BRET signal when in proximity to the GFP. Bottom panel: images of fluorescent signals generated by the different chimeras.

(F) Detection of PTEN-PTEN interaction by BRET in PC3 cells transfected with the indicated plasmids. Bottom panel shows expression levels of the different chimeras. Mean values with associated SD are shown.

(G) Competition assays performed by BRET. Bottom panel, expression levels of the indicated chimeras. Quantification of Un-tagged PTEN cloned in the pcDNA3.1 vector is normalized to β-actin. Mean values with associated SD are shown.

(H) In PC3, BRET signal is collected in a single cell fashion and nuclear emission normalized over average of total emission. Mean values with relative SD are shown.