FIG. 4.

Nuclear translocation of p65 NF-κB in human respiratory epithelial cells in response to stimulation with TNF-α but not to P. aeruginosa DNA. IB3-1 cells in serum-free medium were incubated for different times in control medium or medium with 20 ng/ml TNF-α or 25 μg/ml PAO1 DNA. Cells were fixed and subjected to immunofluorescence analysis with antibodies against the p65 subunit of NF-κB and counterstained with the nuclear stain DAPI. (A) Confocal immunofluorescence microcopy images with cells incubated for 30 min. (B) Quantification of p65 nuclear translocation in cells incubated for 30 min, 2 h, and 6 h. Sample sizes and numbers of nuclei counted: for the 30-min stimulation, n = 139 (control), n = 136 (TNF-α), and n = 118 (PAO1 DNA); for the 2-h stimulation, n = 127 (control), n = 115 (TNF-α), and n = 148 (PAO1 DNA); for the 6-h stimulation, n = 135 (control), n = 113 (TNF-α), and n = 173 (PAO1 DNA).