Figure 2
NF-κB signaling pathway is activated after CSE stimulation in 16HBE cells.
(A) After 15% CSE exposure for different time point, the cells were analyzed by western blot to detect IκBα. β-actin was the loading control. The histograms show the ratio between IκBα and β-actin of 3 experiments. *P < 0.05 vs. 0 h group, #P < 0.05 vs. 1 h group. (B) After 15% CSE exposure for different time point, the cells were analyzed by western blot to detect phosphorylated IκBα (p-IκBα). β-actin was the loading control. The histograms show the ratio between p-IκBα and β-actin of 3 experiments. *P < 0.05 vs. 0 h group, #P < 0.05 vs. 0.5 h group. (C) The nuclear translocation of p65 after CSE exposure in the 16HBE cells. The cells were exposed to15% CSE for the indicated times before fractionation into cytoplasmic and nuclear extracts. β-actin and Lamin B were the loading controls for cytoplasmic and nuclear fractions, respectively. The quantified results can be found as Supplementary Fig. S1. (D) Fluorescence staining of NF-κB-p65. After 6 h of CSE exposure, cells grown on coverslips were subjected to immunofluorescence staining. Scale bar, 50 μm. (E) After exposure to 15% CSE, IL-8 mRNA was measured by RT-PCR at the indicated times. The data were expressed as the means ± SD (n = 3), *P < 0.05 vs. 0 h group, #P < 0.05 vs. 1 h group. (F) The IL-8 concentration was measured by ELISA. The data were expressed as the means ± SD (n = 3), *P < 0.05 vs. 0 h group, #P < 0.05 vs. 1 h group.
