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. Author manuscript; available in PMC: 2013 Jun 1.
Published in final edited form as: Gastroenterology. 2012 Feb 28;142(7):1483–92.e6. doi: 10.1053/j.gastro.2012.02.040

Figure 7.

Figure 7

Pharmacologic CAD inhibitors increase CD-associated NOD2 variant function. (A) Lysates of HEK293T cells transfected with Flag-CAD and HA-tagged NOD2 constructs were immunoprecipitated with Flag antibody followed by immunoblot. (B) NF-κB reporter assay in HEK293T cells cotransfected with control or CAD shRNA plasmids and NOD2 expression constructs. Cells were MDP stimulated (10 ng/mL for 18 hours) and luciferase values normalized to β-galactosidase transfection control values (nLuc). Averages ±SD are shown. (C) Bacterial killing of NOD2 risk variants in HEK293T cells by gentamicin protection assay in the presence or absence of MDP (10 µg/mL). (D) Gentamicin protection assay in HEK293T cells cotransfected with control or CAD short hairpin RNA plasmids and NOD2 expression constructs. (E) Gentamicin protection assay in HEK293T cells transfected with NOD2 constructs pretreated with PALA (250 µmol/L for 1 hour) before infection. Averages ± SD are shown. *P < .05, **P < .01, ***P < .001.