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. Author manuscript; available in PMC: 2014 Nov 10.
Published in final edited form as: Gut. 2012 Jun 8;62(5):695–707. doi: 10.1136/gutjnl-2011-301775

Figure 1. Identification of vimentin as a NOD2 interacting protein.

Figure 1

(a) Schematic showing the structure of vimentin and the vimentin clone identified as binding to NOD2 in our yeast-two-hybrid assay.

(b) Yeast strain AH109 was co-transformed with a plasmid containing the yeast expression vector pGADT7-AD, full-length vimentin cDNA, wild-type NOD2 or CD-associated variant NOD2-R702W as shown. Co-transformants were selected on (Leu-Trp-) plates and spotted onto selective media (His-Leu-Trp-). (Dashed line indicates where irrelevant lanes have been removed).

(c) SW480 cells were transfected with HA-NOD2 or HA-empty vector and extracts immunoprecipitated with HA-11 specific antibodies or non-specific antibody as control. Precipitates were immunoblotted for associated vimentin and NOD2, and direct lysates with HA-11 and vimentin antibodies.

(d) SW480 cells were transfected with HA-NOD2 or HA-empty vector and cell extracts immunoprecipitated with vimentin specific antibodies or non-specific antibody as control. Precipitates were immunoblotted for associated NOD2 and vimentin, and direct lysates with HA-11 and vimentin antibodies.