Acute Pseudomonas challenge in cystic fibrosis mice causes prolonged nuclear factor- κB activation, cytokine secretion, and persistent lung inflammation

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Acute Pseudomonas challenge in cystic fibrosis mice causes prolonged nuclear factor- κB activation, cytokine secretion, and persistent lung inflammation Aicha Saadane, PhD, Jindrich Soltys, DVM, PhD, Melvin Berger, MD, PhD Journal of Allergy and Clinical Immunology Volume 117, Issue 5, Pages 1163-1169 (May 2006) DOI: 10.1016/j.jaci.2006.01.052 Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 Quantitative bacteriology in CF-KO (squares, n = 11) and WT (circles, n = 13) mice inoculated with P aeruginosa. Values represent the mean ± SEMs of log10 of CFU per mouse lung 2, 4, and 6 days after inoculation. The CFUs at day 0 indicate the inoculating dose. Journal of Allergy and Clinical Immunology 2006 117, 1163-1169DOI: (10.1016/j.jaci.2006.01.052) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 Percent PMNs in BAL fluid in CF-KO (squares, n = 26) and WT (circles, n = 25) mice inoculated with P aeruginosa. Results shown are means ± SEMs of the percent PMNs in BAL fluid 2, 4, and 6 days after inoculation. The 0 time point is for unchallenged controls. Journal of Allergy and Clinical Immunology 2006 117, 1163-1169DOI: (10.1016/j.jaci.2006.01.052) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 BAL fluid proinflammatory cytokines in CF-KO (squares, n = 26) and WT (circles, n = 25) mice inoculated with P aeruginosa. Means ± SEMs of concentrations in BAL fluid 2, 4, and 6 days after inoculation. Journal of Allergy and Clinical Immunology 2006 117, 1163-1169DOI: (10.1016/j.jaci.2006.01.052) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 A, Representative EMSA for activated NF-κB in homogenates of perfused whole lungs from CF-KO and WT mice that had undergone BAL and were killed 2, 4, and 6 days after P aeruginosa. B, The amount of complexed NF-κB probe estimated by image scanning and expressed in arbitrary units; CF-KO, closed bars, and WT mice, open bars. Three separate experiments are included. Results shown are means ± SEMs for 3 to 4 mice for each bar. C, Controls for specificity of EMSA: lane 1, labeled probe with extract known to contain active NF-κB; lane 2, labeled probe with no extract; lane 3, labeled probe with extract plus excess of unlabeled NF-κB oligonucleotide; lane 4, labeled probe with extract plus excess of unlabeled AP-1 oligonucleotide. Journal of Allergy and Clinical Immunology 2006 117, 1163-1169DOI: (10.1016/j.jaci.2006.01.052) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 5 A, Western blots showing time course of I-κBα content of homogenate of whole lung from CF-KO versus WT mice inoculated with P aeruginosa. Equal protein application was verified by equal staining for β-actin. Representative of 3 separate experiments. B, Densitometric scan area of total I-κBα on Western blots corrected by normalization to scan area of β-actin. Results are average values of 3 independent experiments. Journal of Allergy and Clinical Immunology 2006 117, 1163-1169DOI: (10.1016/j.jaci.2006.01.052) Copyright © 2006 American Academy of Allergy, Asthma and Immunology Terms and Conditions