Figure 6

TS suppresses RANKL-induced NF-κB activation in RAW 264.7 cells. Confluent RAW 264.7 cells were pre-treated with various concentrations of TS (0.1-2.5 μg/mL) for 2 h followed by RANKL (50 ng/mL) for 30 min. The harvested cells from triplicate tests were subjected to western blot analysis for p-IκB-a. Relative amounts of protein were determined by densitometric analysis (A). RAW 264.7 cells were pretreated with TS (0.1-2.5 μg/mL) overnight, followed by RANKL (50 ng/mL) for 30 min. NF-κB-p65 nuclear translocation analysis were performed by immunofluorescent staining (B). RAW 264.7 cells were incubated with TS (0.1-2.5 μg/mL) for 12 h, followed by the addition of RANKL (50 ng/mL). After 30 min incubation, cells were analyzed for detection of DNA binding of NF-κB by EMSA. The arrowheads indicate the free probe and the specific DNA-probe/transcription factor complex (NF-κB), respectively. Relative NF-κB activity was calculated by densitometric analysis (C). One of three experiments with similar results is shown. Data represent the mean ± SD of three independent experiments. ^### P < 0.001 significantly different from basal. *P < 0.05 and ***P < 0.001 significantly different from RANKL only group.