Fig. 6.
Effect of DHFR knockdown on the expression and phosphorylation of nuclear factor-κB/p65 (NF-κB/p65) and AKT. The expression and phosphorylation levels of NF-κB/p65 and protein kinase B (AKT) were tested by western blot with specific antibodies (dilution 1:500) after DHFR knockdown for 72 h. β-actin served as internal loading control (dilution 1:8,000). (A) Representative gels from 3 experiments are shown. “p-” means phosphorylated protein. p-p65 (s468) and p-p65 (s536) represent NF-κB/p65 phosphorylated at serines 468 and 536, respectively. (B) Densitometric analysis was performed for each protein band relative to β-actin in the same sample using Image LabTM software. To compare the relative quantity (RQ) of proteins between NC-siRNA and DHFR-siRNA groups, the values of density ratio in NC-siRNA group were normalized to 1.00. *vs NC-siRNA, p<0.05; **vs NC-siRNA, p<0.01.