LPS- and LTA-Induced Expression of IL-6 and TNF-α in Neonatal and Adult Blood: Role of MAPKs and NF-κB

Figure 3

Flow-cytometric analysis of p38, ERK1/2, JNK, and NF-κB in monocytes after lipopolysaccharide (LPS) and lipoteichoic acid (LTA) stimulation in vitro. (a) After the stimulation of whole blood (WB) in vitro, the red blood cells were lysed, and the leukocytes were fixed and permeabilized. The leukocytes were stained with modification-specific fluorochrome-conjugated antibodies and an antibody against the CD14 monocyte cell surface marker. The leukocyte population was separated into lymphocytes, granulocytes, and monocytes based on differences in side scatter (granularity) and binding to the CD14 antibody using flow cytometry. The CD14-positive monocyte population was analyzed for the intracellular levels of phosphorylated signaling proteins based on the determination of mean fluorescence intensity (MFI). (b) Measurements of fluorescence intensity from the monocyte subset after staining with the fluorochrome-conjugated antibodies against phosphorylated p38, ERK1/2, JNK, and NF-κB displayed as histograms. The figure shows a fluorescence intensity of unstimulated controls and samples stimulated with LPS and LTA for 15 min.

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