Suppression of osteoclast differentiation and stimulation of osteoblast differentiation by Lrp5 CM and BML284-treated CM. Ob = osteoblasts, CM = osteoblast-derived conditioned medium, A&β = ascorbic acid and β-glycerophosphate, pl = placebo, and Lrp5 = Lrp5 overexpression. The single and double asterisks indicate p < 0.05 and 0.01, respectively. (A) Trap staining of RANKL-stimulated RAW264.7 pre-osteoclasts in response to Ob CM, Lrp5 CM, and BML284-treated CM. (B) Downregulation of NFATc1 and cathepsin K by Lrp5 CM and BML284-treated CM. (C) Enhanced Alizarin-red staining of MC3T3 osteoblasts by Lrp5 CM and BML284-treated CM. (D) Elevation of Lrp5, ALP, osteocalcin, and OPG, and the reduction of RANKL by Lrp5 CM and BML284-treated CM in MC3T3 osteoblasts. (E) Effect of Lrp5 CM on tibial cortical bone. BMD = bone mineral density, N = 10. (F) Downregulation of NFATc1, cathepsin K, and RANKL in the tibia by Lrp5 CM.