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

Figure 4

Effects on phosphorylated p38 MAPK, ERK1/2, JNK, and NF-κB in monocytes after stimulation with lipopolysaccharide (LPS) and lipoteichoic acid (LTA) of whole blood (WB) in vitro. WB was stimulated with LPS (100 ng/mL) or LTA (100 μg/mL) for 15 min. The red blood cells were lysed, and the leukocytes were fixed and permeabilized. The samples were stained with antibodies against CD14 (FITC-conjugated), phosphorylated p38 MAPK (pT180/pY182; Alexa Fluor 647 conjugated), ERK1/2 (pT202/Y204; PE-conjugated), JNK (Thr183/Tyr185; Alexa Fluor 647 conjugated), and/or NF-κB p65 (pS529; PE labeled) and analyzed for the intracellular levels of phosphorylated signaling proteins based on the determination of MFI by flow cytometry. Stimulated samples were compared with unstimulated controls, and the results were expressed as fold change in phosphorylation: , LPS or LTA versus control. , neonatal versus adult.

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