Figure 4
Estrogen protects bone by inducing Fas ligand in osteoblasts to regulate osteoclast survival
Susan A Krum, Gustavo A Miranda-Carboni, Peter V Hauschka, Jason S Carroll, Timothy F Lane, Leonard P Freedman and Myles Brown
- The EMBO Journal (2008) 27, 535 - 545
- doi:10.1038/sj.emboj.7601984
Published online: 24 January 2008
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Fas is expressed in pre-osteoclasts and FasL is expressed in osteoblasts. (A) Bone marrow cultures treated under osteoclastic conditions for 6 days to the pre-osteoclast stage were fixed and immunostained for Fas. The primary antibody was detected with an anti-rabbit IgG Alexa 488 (green fluorescence). DNA was counterstained with DAPI. The cells were then stained for alkaline phosphatase (dark blue cells). The red arrowhead points to clustering of the Fas receptor proteins next to the osteoblast. Parallel cultures were stained with TRAP to identify pre-osteoclasts. (B) Bone marrow cultures treated under osteoclastic conditions for 6 days to the pre-osteoclast stage were fixed and immunostained for FasL and RUNX2. DNA was counterstained with DAPI. (C) A laser scanning cytometer was used to separate cells with low and high RUNX2. The amount of FasL in the presence or absence of 10 nM E2 was quantified in each of these populations.
