p120 Modulates LPS-Induced NF-κB Activation Partially through RhoA in Bronchial Epithelial Cells
Figure 3
p120 downregulation activates NF-κB signaling pathway in LPS-treated bronchial epithelial cells. (a) Cytotoxic effect of LPS on 16HBE 14o-cells was assessed by MTT assay. Data represent the means from three independent experiments and were analyzed by one-way ANOVA test. versuscontrol. (b) The protein expression of p120 was downregulated after being stimulated with LPS (20 μg/mL). (c) Total p120 mRNA was extracted at indicated times. Fluorescent quantitative PCR was performed with SYBR-Green Mastermix PCR system. After comparing the amplifying efficiency with GAPDH, the amplification data were analyzed with the method. The columns represented the relative amplification folds of p120 contrast to GAPDH. Data were expressed as means ± SD (), , and versus control. (d) NF-κB p65 was increased accompanied with IκBα degradation. (e) Nuclear translocation of p65 was detected. Equal amounts of cytoplasmic and nuclear extracts were subjected to Western blot analysis. Lamin B served as a nuclear marker, and β-actin served as a cytoplasmic marker. (f, g) IL-8 production was increased. (f) Supernatants were collected and assayed for IL-8 by ELISA. Data were expressed as means ± SD (), versus control group (0 min). (g) IL-8 mRNA was measured by fluorescent quantitative real-time PCR. The area at 0 min was assigned as 1.0. Data were expressed as means ± SD (), , and versus control group (0 min).
