Fig. 5. B7–H3 deficiency-mediated type-I IFN-dependent osteoclastogenesis inhibition is independent of c-Fos, iNOS, and SOCS1.

A, B M-CSF (40 ng/ml) was added to monocyte culture for three days. OCPs that were transiently transfected with B7–H3-specific siRNA or control siRNA (40 nM) were induced to differentiate using M-CSF (40 ng/ml) and RANKL (80 ng/ml) for seven days. The expression of B7–H3, c-Fos and iNOS protein during osteoclast differentiation was detected via Western blot. The bar graphs show the densitometry analyses of the gels depicted relative to the internal control at day 7 of differentiation. B, C OCPs were transiently transfected with B7–H3-specific siRNA or control siRNA (40 nM) for one day. The expression of SOCS1 and iNOS mRNA was analyzed using RT-qPCR. The mRNA levels were normalized relative to GAPDH expression.