Figure 4. Effects of cold-inducible RNA-binding protein (CIRBP) on NF-κB signaling pathway. Cells were transfected with pcDNA3.1, pc-CIRBP, siNC or siCIRBP and cultured in serum-free medium with hypoxia. Protein expression was evaluated by western blot analysis. The band intensity was assessed by Image Lab™ software. The phosphorylation rate is reported as the relative intensity of phosphorylated kinases/total kinases and the final results were normalized by GAPDH. pc-CIRBP: pcDNA3.1 containing CIRBP coding sequence; siCIRBP: CIRBP-specific small interfering RNA; siNC: siCIRBP negative control; IκBα: inhibitor of nuclear factor κBα; p-IκBα: phosphorylated IκBα; p-p65: phosphorylated p65. Data are reported as means±SD of 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001, pc-CIRPB compared to pcDNA3.1 and siCIRBP compared to siNC (ANOVA).