MassIVE MSV000086418

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Fam20C Dual Regulation of Bone Resorption and Breast Cancer Bone Metastasis through Osteopontin and BMP4

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Description

Samples: Conditioned medium from cultured wild-type or Fam20C-VavCre bone marrow cells. Six 10-cm plates of bone marrow cells from each genotype (Wild-type or Fam20C-VavCre) of 2-month-old female littermate mice at a density of 2.8*107 cells/plate were cultured in the presence of 40 ng/mL of mouse M-CSF for 3 days. Cells were washed with PBS for four times and with serum-free medium twice. Then cells were cultured in serum-free medium (MEMalpha without phenol red) for 48 hours. Conditioned medium was collected and spin down at 1,000 g for 5 minutes. The supernatant was spin down at 10,000 g for 20 minutes. The supernatant was added with 1 mM Na3VO4, 1 mM PMSF and 1x protease inhibitor cocktail (Sigma-Aldrich), and concentrated with Amicon Ultracel-3K units (EMD Millipore). Protein concentrations were measured with Pierce BCA protein assay (Fisher Scientific), and 600 ug of protein each sample were subjected to following procedures [doi:10.25345/C5S783] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Fam20C, breast cancer, osteoclast, bone resorption, bone metastasis

Contact

Principal Investigators:
(in alphabetical order) 		Yihong Wan, UT Southwestern Medical Center, United States
Submitting User: mogoodarzi
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