Figure 6:

Low-magnitude CS switch attenuates inflammatory signaling activated by LPS and high-magnitude CS. Pulmonary ECs were primed by LPS for 15 min before exposure to 6-h 18% CS or 3-h 4% CS followed by 3-h 18% CS. (A) 18% CS-stimulated phosphorylation of p-NFκB and expression of ICAM1, VCAM1, and E-selectin. These effects were reduced in ECs preconditioned at 4% CS before 18% CS. (B) Intracellular localization of NFκB was detected by immunofluorescence imaging. ECs were exposed to 4% CS or to 4% CS before or after 18% CS. Bar = 10 µm. (C) Western blot analysis of ICAM1 expression. Inclusion of 4% CS before or after 18% CS reduced ICAM1 expression in pulmonary ECs activated by LPS and 18% CS. Membrane reprobing for α-tubulin was used as a normalization control. Bar graphs represent analysis of Western blot data; *, p < 0.05 vs. 18% CS.