Figure 4

Blimp1 mediates the stimulatory effects of TG2 knockdown on osteoclastogenesis. (a) BMMs were transfected with TG2 and/or Blimp1 siRNA and cultured for two days with M-CSF (30 ng/ml) and RANKL (100 ng/ml). The mRNA levels of TG2 and Blimp1 were analyzed by real-time PCR. **P < 0.005 between indicated groups. (b,c) siRNA-transfected cells were cultured for three days with M-CSF and RANKL. The cultured cells were fixed and subjected to TRAP staining. Scale bar, 100 μm (b). TRAP-positive MNCs were counted, and the surface area per osteoclast was measured. *P < 0.05; **P < 0.005 between indicated groups (c). CTL, control. (d) siRNA-transfected cells were cultured for two days with M-CSF and RANKL. Cell lysates were prepared and subjected to Western blotting. C, control siRNA; T, TG2 siRNA; B, Blimp1 siRNA. Full length Western blots are presented in Supplementary Figure 10.