Fig. 3
From: mTORC1 impedes osteoclast differentiation via calcineurin and NFATc1
Late mTORC1 deletion in myeloid lineage enhances osteoclastogenesis. Analyses of Raptorfl/fl-Lyz-Cre cKO mice and littermate controls. a, b Expression of osteoclast markers (a) and osteoclastogenic transcription factors (b) in bone marrow osteoclast differentiation cultures (n = 6). c Representative images of TRAP-stained osteoclast differentiation cultures. d Osteoclast resorptive activity measured by calcium release from bone plates (n = 16). e Osteoclast precursor proliferation by BrdU incorporation (n = 10). f, g mTORC1 signaling in bone marrow osteoclast differentiation cultures from Raptorfl/fl-Lyz mice or littermate controls 50 h after RANKL treatment. f Phosphorylation of mTOR; g Phosphorylation of S6K1. p/T, ratio of phosphorylated/total protein. h–t Bone phenotype in 3-month-old male mice. h Serum CTX-1 bone resorption marker (n = 8). i Serum P1NP bone formation marker (n = 8). j Representative μCT images of the trabecular bone of the tibial metaphysis (top) and the entire proximal tibia (bottom). k–p Quantification of trabecular bone volume and architecture in proximal tibiae by μCT (n = 8–10). k BV/TV bone volume/tissue volume ratio; l BS bone surface; m Tb.N trabecular number; n Conn. D. connectivity density; o Tb.Sp trabecular separation; p SMI structure model index. q–t Bone histomorphometry of distal femurs (n = 6). Oc.S osteoclast surface, B.S bone surface, Oc.N osteoclast number, Ob.S osteoblast surface, Ob.N osteoblast number. Error bars, SEM; *, p < 0.05; **, p < 0.01; ***, p < 0.005; ****, p < 0.001; n.s. non-significant. R raptor. Full-size scans of immunoblots are shown in Supplementary Fig. 7
