FIGURE 1.

Gene silencing and function of DGCR8, Dicer1 or Ago2 in osteoclast precursors. Quantitative real time PCR was performed with primer for mouse DGCR8 (A), Dicer1 (B), or Ago2 (C). The data represent the means ± S.D. of three experiments in triplicate. The loss of DGCR8 (D), Dicer1 (E), or Ago2 (F) protein expression levels by siRNA gene silencing was analyzed by immunoblotting (IB) using nuclear or whole cell extracts. Quantitative image analysis of these protein expression levels was normalized to GAPDH (% scr). scr, scrambled. miRNA RT-PCR (G and H) and quantitative real time RT-PCR (I) were performed to measure miRNA-223 and miRNA-155 expression levels. The PCR products were confirmed using a 20% polyacrylamide gel, with U6 small nuclear RNA as a loading control (G and H). PCR products were normalized to U6 level for each reaction (I). The data represent the means ± S.D. of three experiments in triplicate.