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Blood, Vol. 109, Issue 8, 3253-3259, April 15, 2007
MafB negatively regulates RANKL-mediated osteoclast differentiation
Blood Kim et al.
109: 3253
Supplemental materials for: Kim et al, Vol 109, Issue 8, 3253-3259
Materials and methods Phagocytosis assay
BMMs were infected with retroviral supernatant for 8 hours and cells were cultured for 2 days with M-CSF (30 ng/mL) or M-CSF (30 ng/mL) and RANKL (100 ng/mL). Fluorescein-conjugated zymosan A (Saccharomyces cerevisiae) Bio Particle (Molecular Probes, Eugene, OR) was added to cultured cells in 96-well culture plates (20 µg/0.2 mL/well). After 1 hour of incubation, cells were washed with PBS to remove the particles that were not incorporated by the cells. Cells were fixed and observed with UV illumination under the microscope. MTT assay
The cultured cells were incubated with 25 µL/well of MTT solution (5 mg/mL) for 2 hours at 37°C. After incubation for 24 hours, formazan crystals were dissolved in 100 µL of extraction buffer (20% SDS/50% DMF) and the absorbance was measured at 570 nm using a microplate reader.
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