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. 2008 Nov 27;18(4):737–752. doi: 10.1093/hmg/ddn404

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Figure 9. — Expanded polyglutamine proteins interact with mitofusin proteins. Bacterial expressed GST, GST-HttQ28 and GST-HttQ74 fusion proteins were purified by GS-agarose affinity chromatography. Next extracts were prepared from HeLa cells transfected with the myc-tagged Mfn2 expression construct, which were then passed over the GS-agarose columns, repeatedly. After extensive washing, the proteins that bound to the column were analyzed by immunoblotting. Upper panel: proteins immunoblotted with an anti-myc antibody reveal that the myc-tagged mfn2 protein binds preferentially to GST-HttQ74 fusion protein, but not GST alone or GST-HttQ28 fusion protein. Middle panel: proteins immunoblotted with an anti-Mfn2 antibody reveal preferential binding of both endogenous (arrow head) and exogenous myc-tagged (asterisk) Mfn2 proteins with GST-HttQ74 protein. Lower panel: immunoblot of the lysates with a anti-GST antibody detects the appropriate size GST proteins used for the bindings assays.