Figure 4.

Saikosaponin D attenuated RANKL-induced activation of NF-κB pathway. (A) SSD suppressed RANKL-induced relative phosphorylation of P65 and IκBα and the degradation of IκBα. BMMs were treated without or with various concentrations of SSD (0.5, 1 and 2 μmol/L) for 1 h and then were treated with RANKL (50 ng/mL) for 15 min. Total proteins extracted from BMMs were probed for protein levels using immunoblot analyses. (B) Expressions of IκBα and p‐IκBα relative to β‐actin and p‐NFκB P65 relative to total NF-κB P65. (C) Nuclear translocation of P65 indicated by immunofluorescence. (D) The relative frequency of cytoplasmic and nuclear P65 in each experimental group was quantified using Image J. Original magnification, ×200. Scale bar, 200 μm. Mean ± SD is used to describe the data. Above experiments were conducted independently at least 3 times. ****P <0.0001, ***P<0.001.

Abbreviation: ns, not significant.