Portland Press

Figure 3

$(A) Osteoblasts were pretreated with TLR2- or TLR4-siRNA for 24 h, followed by the addition of rhHMGB1 (150 μg/l) to the culture medium for 24 h. Cells were harvested and fractionated into the cytoplasm and the nucleus. Lysates were then separated on a 10% SDS/PAGE and subjected to western blotting with anti-p65. GAPDH and Lamin B1 were used as markers for the cytoplasmic and nuclear fractions. (B) The NF-κB p65 protein levels that were translocated to the nucleus were quantified by densitometric analyses. One-way ANOVA was performed to determine statistical significance. Pretreatment with TLR2- or TLR4-siRNA decreased nuclear NF-κB p65 subunit levels, whereas rhHMGB1 enhanced it (*P<0.05).$

Effects of rhHMGB1 and TLR2- or TLR4-siRNA on NF-κB expression in osteoblasts

(A) Osteoblasts were pretreated with TLR2- or TLR4-siRNA for 24 h, followed by the addition of rhHMGB1 (150 μg/l) to the culture medium for 24 h. Cells were harvested and fractionated into the cytoplasm and the nucleus. Lysates were then separated on a 10% SDS/PAGE and subjected to western blotting with anti-p65. GAPDH and Lamin B1 were used as markers for the cytoplasmic and nuclear fractions. (B) The NF-κB p65 protein levels that were translocated to the nucleus were quantified by densitometric analyses. One-way ANOVA was performed to determine statistical significance. Pretreatment with TLR2- or TLR4-siRNA decreased nuclear NF-κB p65 subunit levels, whereas rhHMGB1 enhanced it (*P<0.05).

This Feature Is Available To Subscribers Only