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. Author manuscript; available in PMC: 2011 Jun 1.
Published in final edited form as: J Cell Biochem. 2010 Jun 1;110(3):645–659. doi: 10.1002/jcb.22575

Figure 6. Activation of NF-κB by S100A12 ligation of RAGE is inhibited by mAbGB3.1 and sRAGE.

Figure 6

Stable transfectants of HeLa cells expressing full length and cytoplasmic tail deleted (signaling deficient) RAGE were generated as described. Expression of RAGE in cells was confirmed by A. Western blotting using anti-RAGE, Lane 1, untransfected HeLa cells; Lane 2, Full length RAGE transfectants and Lane 3, Tail deleted RAGE transfectants and by B. FACS analysis. Unstained cells are represented in the background. Low-level endogenous expression is seen in untransfected cells. C. Activation of NF-κB by S100A12 ligation of RAGE is inhibited by mAbGB3.1 and sRAGE: HeLa cells expressing full length or tail deleted RAGE were transiently transfected with an NF-κB-responsive cis-reporter gene and a pSV-β-Galactosidase control vector. S100A12 activation in the presence and absence of inhibitors and analysis of enzymes in lysates was done as described. The ratio of Luc/Gal served to normalize for Luc activity. Luc/Gal ratio in transfected unactivated control, which accounted for endogenous activity, was considered 100%. Activity in tail deleted RAGE accounted for RAGE-unrelated background. All values represent the mean ± S.D. (n = 2).