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. 2015 Nov 16;212(12):1993–2002. doi: 10.1084/jem.20151069

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© 2015 LaFlam et al.

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Figure 2. — ACNS1 is an NF-κB-responsive element. (A) Comparison of consensus ACNS1 sequence to κB sequence motif. (B) EMSA using nuclear lysates from 293T cells transfected with p52-FLAG, incubated with biotinylated ACNS1 probe. Untagged WT and mutant probes (first, second, or both κB sites, respectively), and anti-FLAG antibody included in specified lanes. (C) Relative luminescence after chemiluminescent detection of β-gal in cellular lysates of 293T cells 48 h after transfection with TK-β-gal or CNS1-TK-β-gal plasmid ± RelB-FLAG and p52-FLAG. (D) Relative luminescence after chemiluminescent detection of β-gal in cellular lysates of 293T cells, 48 h after transfection with RelB-FLAG and p52-FLAG and WT or mutated CNS1-TK-β-gal. In C and D, normalized for transfection efficiency using pRL-CMV. All data are representative of at least three independent experiments. Statistical analysis using Student’s t test: ns, not significant; *, P < 0.05; **, P < 0.01; ****, P < 0.0001.