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. 2018 Oct 31;9:2366. doi: 10.3389/fimmu.2018.02366

Figure 3.

Figure 3

R70W CARD9 inhibits NF-κB transcriptional activity. (A, C) Schematic summaries of the experimental conditions in (B,D) respectively, adjusted from (13). (B, D) MALT1-deficient HEK293T cells were transfected with WT, R70W, L213LI and R70W/L213LI CARD9 as indicated, with fixed total DNA quantities, with or without MALT1 as indicated, together with an NF-κB-dependent luciferase reporter expression plasmid and a constitutively expressed β-galactosidase reporter gene. Luciferase values were normalized against β-galactosidase and expressed as fold induction compared to WT CARD9 without MALT1. Expression of MALT1 and CARD9 protein measured by western blot are shown below the graphs. (E) WT HEK293T cells were co-transfected with decreasing doses of L213LI DNA and increasing doses of R70W/L213LI CARD9 DNA (ng/well, doses specified under graph). The total DNA amount was kept constant at 400ng/well. Luciferase values were expressed as fold induction compared to L213LI CARD9 alone. All values under the dotted line are significant.Results shown in (B,D,E) are mean +/- standard deviation of 4 replicates. One out of 2–3 representative experiments is shown. Statistical analysis was performed on reporter assay data with one-way ANOVA and Tukey's multiple comparison's (B, D, E) post-testing. The most relevant statistical differences are shown, a list of all p-values is provided in Supplemental Tables 2–4. p < 0.001 (***) in all panels, in (B,D) only for reporter assays with MALT1 reconstitution.