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. Author manuscript; available in PMC: 2011 Nov 1.
Published in final edited form as: Gastroenterology. 2010 Jul 14;139(5):1630–1641.e2. doi: 10.1053/j.gastro.2010.07.006

Figure 4.

Figure 4

CD-associated NOD2 mutants are impaired in signaling, anti-bacterial killing, and autophagy. (A) NF-κB luciferase reporter assay in HEK293T cells transfected with Nod2 plasmids and stimulated with MDP (1µg/mL) for 18h. Luciferase values normalized to β-gal transfection control (nLuc), mean ±SD. (B) Gentamycin protection assay of HEK293T cells transfected with Nod2 plasmids, mean ±SD. (C) NF-κB luciferase reporter assay in HEK293T cells transfected with Nod2 and IKK dominant negative (DN) constructs, mean ±SD. (D) Gentamycin protection assay of HEK293T cells transfected with Nod2 and IKK DN plasmids, mean ±SD. (E) Autophagosomal maturation assessed by flow cytometric analysis of GFP-mCherry-LC3 in HEK293T cells transfected with vector, Nod2 or Nod2 L1007fs plasmids. Histogram of GFP MFI of the mCherry-positive population (left). Quantification of GFP/mCherry MFI ratio normalized to vector transfected sample, mean ±SD (right, n=7). (F) LC3 immunoblots of HEK293T cells transfected with Nod2 or Nod2 L1007fs plasmids treated with MDP (1µg/mL) or rapamycin (25µg/mL) for the indicated times (hours). Quantification of LC3-II amount indicated by LC3-II/tubulin ratio (right).

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