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. Author manuscript; available in PMC: 2011 Jun 1.
Published in final edited form as: Int J Biochem Cell Biol. 2010 Mar 27;42(6):1039–1045. doi: 10.1016/j.biocel.2010.03.014

Fig.5. AR inhibition prevents LPS-induced phosphorylation of CREB and PKA in macrophages.

Fig.5

A) Growth-arrested macrophages were pre-incubated with and without AR inhibitors (2 μM fidarestat) for 24 h followed by the incubation with LPS (1 μg/ml) for 0, 5, 15, 30, 45 and 60 min. The whole cell lysates were subjected to Western blot analysis and probed with p-CREB, CREB, p-PKA, PKA and GAPDH antibodies. Fold change was calculated based on the densitometric analysis using Kodak Image station. Data shown are representative of three independent experiments. B) RAW264.7 cells were transfected with PathDetect® reporter plasmid pCRE-hrGFP as described in methods and treated with and without 2 μM fidarestat or 50 μg/ml H-89 (PKA inhibitor) for an hour prior to treatment with LPS 1μg/ml for 24 h. Ethanol-fixed cells were washed twice in cold PBS and analyzed for GFP-positive cells using FACSCanto. Percentage of positive cells was plotted. Experiment was carried with triplicates. All the data are expressed as percentage of Mean ± SEM (N = 4). *P < 0.001 as compared to pCRE control cells, #P < 0.001 compared to LPS+pCRE-treated cells