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. Author manuscript; available in PMC: 2009 Mar 1.
Published in final edited form as: Surgery. 2007 Dec 27;143(3):334–342. doi: 10.1016/j.surg.2007.09.039

Fig. 4. Exogenous ghrelin inhibits LPS-induced translocation of p65 subunit of NFκB into the nuclear compartment in murine RAW 264.7 macrophages.

Fig. 4

Fig. 4a, Western analysis; RAW 264.7 macrophages were pre-treated with various agents including ghrelin (0-100nM), D[lys-3]-GHRP-6 (1μM) and a selective NFκB inhibitor (6-Amino-4-(4-phenoxyphenylethylamino)quinazoline) (1μM) 60 minutes prior to the LPS (1μg/ml) treatment, nuclear extracts were prepared as described in the Materials and Methods section and were subjected to Western analysis for assessment of nuclear content of p65 NFκB; lane a, no treatment; lane b, ghrelin; lane c, LPS (1μg/ml); lane d, ghrelin (1nM) + LPS (1μg/ml); lane e, ghrelin (10nM) + LPS (1μg/ml); lane f, ghrelin (100nM) + LPS (1μg/ml); lane g, D [lys-3]-GHRP-6 (1μM) +ghrelin (10nM) + LPS (1μg/ml); lane h, NFκB inhibitor + ghrelin (10nM) + LPS (1μg/ml). Fig. 4b, Immuno-florescent staining for p65 subunit of NFκB; RAW 264.7 cells were pre-treated with various substances 60 min prior to stimulation with LPS for 15 min; cells were stained with DAPI and p65 subunit of NFκB antibody; A, light microscopic image of the cells; B, no treatment; C, LPS treatment- the p65 subunit translocates to nuclear region; D, LPS(1μg/ml)+ ghrelin 10nM; E, LPS(1μg/ml)+D[lys-3]-GHRP-6 pre-treatment + ghrelin; F, LPS(1μg/ml)+ D[lys-3]-GHRP-6 pre-treatment + ghrelin + NFκB inhibitor.