Research Paper Volume 15, Issue 3 pp 718—733

Figure 7. Role of Raptor in the effect of IL-6 on promoting osteogenic differentiation inf MEFs. (A) Effect of BMP9, IL-6, and/or IL-6 knockdown on p-Stat-3 and Sta3-3 was measured using western blot assay. (B) Quantification of western blot assay show the effect of BMP9, IL-6, and/or IL-6 knockdown on p-Stat-3 and Sta3-3. (C) Effect of IL-6 and/or Stat-3 inhibition on Raptor protein was measured using western blot. (D) Effect of IL-6 and/or Stat-3 inhibition on Raptor mRNA was measured using real-time PCR. (E) Quantification of western blot assay show the effect of IL-6 and/or Stat-3 inhibition on Raptor protein level. (F) Effect of IL-6, Stat-3 inhibition, and/or Raptor knockdown on Runx2 protein was measured using western blot. (G) Effect of IL-6, Stat-3 inhibition, and/or Raptor knockdown on ALP activity was measured using histochemical stain. (H) Quantification of histochemical stain show the effect of IL-6, Stat-3 inhibition, and/or Raptor knockdown on ALP activity. (I) Regulation of p-Stat-3 on Raptor promoter activity was measured using ChIP assay (P1: primer 1, P2: primer 2, P3: primer 3). (^* P < 0.05, ^** P < 0.01; HO-3867, Stat-3 specific inhibitor).