Figure 2: Inhibition of NFAT activation in osteoblasts increases osteoblast differentiation and bone formation. (a) Primary osteoblasts were harvested from the calvariae of control and dnNFATOB mice and cultured for 7 (alkaline phosphatase, ALP) or 21 (von Kossa) days in the presence of -glycerophosphate and ascorbic acid-2-phosphate to induce osteoblast differentiation. Cells were stained for ALP activity (red) or for mineralization by von Kossa (black). Images are representative of three independent experiments, each repeated in duplicate. (b) Femora from 12-week-old control and dnNFATOB mice were stained with Goldner’s Trichrome (blue) and von Kossa (black) staining to show mineralized bone. Magnification, 200x. (c–f) Histomorphometrical indices were measured from 12-week-old control () and dnNFATOB () mice and show (c) bone volume/tissue volume (BV/TV), (d) mineral apposition rate (MAR) (μm/day), (e) osteoblast number/bone surface (N.Ob/BS), and (f) osteoclast number/bone surface (N.Oc/BS). Values represent individual mouse measurements with the mean indicated by the black bar; * when compared to control mice.