Fig. 4

Effects of ciprofloxacin and levofloxacin on NF-κB activation in unstimulated and LPS-stimulated microglia. Cells were subcultured for 24 h in 10% serum-containing medium, which was replaced with serum-free medium before stimulation with ciprofloxacin (CPFX) and levofloxacin (LVFX) (500 μg/ml) ± 100 ng/ml LPS. a Cells were processed for NF-κB p65 immunostaining. Experiments were performed three times and representative confocal images showing subcellular localization of p65 are shown. Scale bars, 10 μm. b Fluorescence intensity profiles along an ideal oriented line across a representative cell (yellow arrows in the third line of (a)). c The fluorescence intensity of cytoplasmic and nuclear p65 subunit was calculated using ImageJ software and results are presented as a percentage of nuclear (gray bars) over cytoplasmic NF-κB p65 (white bars). Data are mean ± SEM from five random fields of three separate experiments