ld mounting medium. The samples were observed on an LSM 510 Meta confocal microscope, and data were processed using the software provided by the manufacturer or Image J software. HEPES, 0.2 mM EDTA, 25% glycerol, 1 mM dithiothreitol, 0.5 mM phenylmethylsulfonyl fluoride, and complete protease inhibitors. 11885967” Protein concentrations were determined by Bio-Rad protein assay kit. The EMSA probes were 21278085” double-stranded oligonucleotides containing a murine IL-6 C/ EBP binding site, or a NF-kB consensus oligonucleotide. C/EBP probes were labeled with a ATP. NF-kB probes were labeled with c ATP. DNA binding reactions were performed at room temperature in a 25 ml reaction mixture containing 6 ml of nuclear extract and 5 ml of 56 binding buffer Ficoll, 50 mM HEPES pH 7.9, 5 mM EDTA, 5 mM dithiothreitol). The remainder of the reaction mixture contained KCl at a final concentration of 50 mM, Nonidet P-40 at a final concentration of 0.1%, 1 mg of poly, 200 pg of probe, bromphenol blue at a final concentration of 0.06%, and water to final volume of 25 ml. Samples were electrophoresed through 5.5% polyacrylamide gels in 16 TBE at 190 V for approximately 3.5 h, dried under vacuum, and exposed to X-ray film. For supershifts, nuclear extracts were preincubated with antibodies for 0.5 h at 4uC prior to the binding reaction. The following antibodies were purchased from Santa Cruz, CA: NF-kB p50, p52, p65, RelB, cRel, C/EBPa, C/EBPb, C/EBPd, C/EBPe, C/EBPc, and normal rabbit immunoglobulin G. Statistical Analysis All values were expressed as the mean 6 S. E. M. Significance was assigned where p,0.05. Data sets were analyzed using Student’s t test or one-way ANOVA, with individual group means being compared with the Student-Newman-Keuls multiple comparison test. Acknowledgments We greatly appreciate the gift of the expression vectors for C/EBPb, IL-6 promoter-luciferase construct containing a mutated NF-kB binding site provided by Richard C. Schwartz, IL-6 promoter-luciferase construct containing a mutated C/EBP binding site provided by Gail A. Bishop, and 2XC/EBP-luc reporter plasmid provided by Peter Johnson. Electrophoretic Mobility Shift Assay Nuclear extracts of MLE12 cells were prepared as follows. Cells were lysed in 15 mM KCl, 10 mM HEPES, 2 mM MgCl2, 0.1 mM EDTA, 1 mM dithiothreitol, 0.1% Nonidet P-40, 0.5 mM phenylmethylsulfonyl fluoride, and complete protease inhibitors for 10 min on ice. Nuclei were pelleted by centrifugation at 14,0006 g for 20 sec at 4uC. Proteins were extracted from nuclei by incubation at 4uC with vigorous vortexing in buffer C Alveolar epithelial type II cell: defender of the alveolus revisited. Respir Res 2: 3346. Whitsett JA, Wert SE, Weaver TE Alveolar surfactant homeostasis and the pathogenesis of pulmonary disease. Annu Rev Med 
61: 105119. Mason RJ Biology of alveolar type II cells. Respirology 11 Suppl: S1215. Kannan S, Huang H, 345627-80-7 site Seeger D, Audet A, Chen Y, et al. Alveolar epithelial type II cells activate alveolar macrophages and mitigate P. Aeruginosa infection. PLoS One 4: e4891. 5. Vanderbilt JN, Mager EM, Allen L, Sawa T, Wiener-Kronish J, et al. CXC chemokines and their receptors are expressed in type II cells and upregulated following lung injury. Am J Respir Cell Mol Biol 29: 661668. Thorley AJ, Goldstraw P, Young A, Tetley TD Primary human alveolar type II epithelial cell CCL20 induced dendritic cell migration. Am J Respir Cell Mol Biol 32: 262267. Sato K, Tomioka H, Shimizu T, Gonda T, Ota F, et al. Type II alveolar