Figure 5

TG2 overexpression inhibits osteoclast formation and Blimp1 expression in RAW264.7 cells. TG2-overexpressing RAW264.7 cells were generated by lentiviral transduction. (a) TG2 mRNA levels in the control and TG2-overexpressing cells were measured by real-time PCR (left panel). **P < 0.005 versus control stable RAW264.7 cells. Whole cell lysates were subjected to Western blot analysis with anti-TG2 antibody (right panel). (b) The control and TG2-overexpressing RAW264.7 cells were cultured with RANKL for four days, and TRAP staining was performed. Red arrows indicate MNCs. Blue arrowheads indicate mononucleated cells. Scale bar, 100 μm. (c) TRAP-positive MNCs were counted, and the surface area per osteoclast was measured. **P < 0.005 versus control stable RAW264.7 cells. (d) The mRNA levels were measured by real-time PCR with normalization to the level of HPRT. *P < 0.05; **P < 0.005 versus control stable RAW264.7 cells. (e) Whole cell lysates were subjected to Western blot analysis using the indicated antibodies. Anti-V5 antibody was used to detect ectopic TG2. Full length Western blots are presented in Supplementary Figure 10.