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To perform immunoblotting assays, cells were seeded in clear 6-well plates (Corning Costar, Fisher Scientific, USA) at a density of 300,000 cells/well, while for immunofluorescence assays cells were seeded at a density of 20,000 cells/well in covers placed in 24-well plates.
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When the bone is entirely covered, place it somewhere safe to let it dry.
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The trays were covered and placed into plastic bags to prevent evaporation.
For LSCM imaging, cells were seeded onto a glass cover slip placed in six well plates.
Cells were cultured on sterile round cover slips placed in 24-well plates.
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