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. Author manuscript; available in PMC: 2012 Jun 19.
Published in final edited form as: Nat Genet. 2011 Oct 9;43(11):1066–1073. doi: 10.1038/ng.952

Figure 4. Functional analyses of NOD2 variants.

Figure 4

HEK293T cells were transfected with NOD2 constructs and fixed using paraformaldehyde 4% at 24h post transfection. Cells were then subjected to immunofluorescent staining to detect NOD2 and fluorescence was collected using a confocal microscope. Image gallery displays a single confocal section.

HEK293T cells were transfected with NOD2 constructs and reporter plasmids encoding firefly luciferase cloned under a promoter containing NF-kB elements and with a plasmid encoding renilla luciferase as a transfection control. After 24h, cells were then stimulated with MDP-LL or MDP-LD (1ug/ml) for 6h. Transcriptional activation was quantified by ratios of firefly luciferase activity to renilla luciferase activity. Data were normalized to the unstimulated condition with empty vector transfection. Statistical analyses were performed using Student t-test. (* p<0.05). Cell lysates were also collected and subjected to western blot analysis to detect NOD2 and actin expression levels.