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Blood, Vol. 107, Issue 8, 3098-3105, April 15, 2006
Thalidomide derivative CC-4047 inhibits osteoclast formation by down-regulation of PU.1
Blood Anderson et al.
107: 3098
Supplemental materials for: Anderson et al
Files in this Data Supplement:
- Figure S1A-B (PDF, 16.8 KB) -
After an incubation period of 21 days cell survival was determined by Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining according to the manufacturer’s instructions (Bender MedSystems, Vienna, Austria) and analyzed by flow cytometry using a FACS Caliber flow cytometer . The percentage of viable cells, negative for both, Annexin V-FITC and PI, was determined.
- Figure S1C-D (PDF, 11.8 KB) -
Cells from OCL assays were detached using cell dissociation buffer (Sigma-Aldrich, St. Louis, MO, USA) and stained with anti-CD116 (Serotec, Oxford, UK). Data were acquired on a Dako-Cytomation MoFlo cytometer. Dyes were excited at 520 nM with mixed gas green laser to minimize autofluorescence of the cultured cells. Cell doublets and clusters were gated from the analysis using doublet discrimination (forward scatter pulse height by pulse width). Events with low forward scatter (dead cells and debris) were gated from the analysis.
- Figure S1E-F (PDF, 11.7 KB) -
Cells from OCL assays were detached using cell dissociation buffer (Sigma-Aldrich, St. Louis, MO, USA) and stained with anti-CD114 (Serotec, Oxford, UK). Data were acquired on a Dako-Cytomation MoFlo cytometer. Dyes were excited at 520 nM with mixed gas green laser to minimize autofluorescence of the cultured cells. Cell doublets and clusters were gated from the analysis using doublet discrimination (forward scatter pulse height by pulse width). Events with low forward scatter (dead cells and debris) were gated from the analysis.
- Figure S1G-H (PDF, 19.6 KB) -
Cells from OCL assays were detached using cell dissociation buffer (Sigma-Aldrich, St. Louis, MO, USA) were stained with anti-CD34 (BD Bioscience, San Jose, CA). Data were acquired on a Dako-Cytomation MoFlo cytometer. Dyes were excited at 520 nM with mixed gas green laser to minimize autofluorescence of the cultured cells. Cell doublets and clusters were gated from the analysis using doublet discrimination (forward scatter pulse height by pulse width). Events with low forward scatter (dead cells and debris) were gated from the analysis.
- Figure S1I (PDF, 10.3 KB)
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