Figure 5

Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting results in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. (A) Changes in the expression of Cebpb, members of the Runx family, and SRY (sex determining region Y)-box 2 (Sox2) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(negative control) + 925) shows slightly reduced expression of Sox2. Runx2 knockdown (N + 1623) shows increased expression of Sox2 and Runx1, but decreased expression of other Runx2 isoforms and Runx3. Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) was also performed. (B) Western blotting results. N indicates negative control stealth siRNA, 925 and 1623 indicate Cebpb and Runx2 type1 siRNA, respectively (final concentration 10 nM). (C) Changes in the expression of adhesion molecules, epithelial-mesenchymal transition (EMT) markers, and mesenchymal markers in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Cebpb knockdown (N(Negative control) + 925) caused increased expression of EMT markers snail family zinc finger 2 (Snai2) and decreased expression of the mesenchymal marker vimentin (Vim) and the epithelial marker E-cadherin (Cdh1). Runx2 type1 knockdown (N + 1623) caused decreased expression of adhesion molecules Cdh1 and integrin alpha 6 (Itga6), decreased expression of Vim, and increased expression of Snai2. Concomitant Cebpb and Runx2 type1 knockdown (925 + 1623) caused decreased expression of Cdh1, Itga6 and VIm, and increased expression of mesenchymal marker N-cadherin (Cdh2) and Snai2. (D) Changes in biglycan (Bgn), decorin (Dcn), bone morphogenic proteins (Bmps) in mHAT9d cells 48 h after transfection with Cebpb and Runx2 type1 stealth siRNA. Concomitant Cebpb and Runx2 type1 siRNA knockdown (925 + 1623) shows a markedly increased expression of Bgn and Dcn (encoding two small leucine-rich peptidogycans), and increased expression of Bmp4/6.