Figure 10G-H. Effect of MSDK on RUNX2, β1 integrin, NFκB and metabolic proteins. Metabolic proteins such as PPARγ, GLUT4 and Insulin Rβ expression were also measured in (G) osteoblasts grown in transwell co-cultures and (H) in osteoblasts and osteoclasts grown in layered co-cultures. Cell lysates were prepared on day 21 from the bottom (osteoblasts) and top (osteoclasts) chambers in the transwell co-culture and from the whole plate (both osteoblast and osteoclast) in the layered co-culture. Protein levels were normalized against β-actin and then to Os-/MSDK. Mean protein levels were analyzed and compared between groups (Os-/MSDK, Os+/Veh, Os+/MSDK). *=p<.05, **=p<.01 and **=p<.001; One-way ANOVA followed by Bonferroni’s post-hoc multiple comparison t-test (n=6 per group).