Copyright © 2009 American Medical Association. All rights reserved. From: 7β-Hydroxycholesterol Induces Apoptosis and Regulates Cyclooxygenase 2 in Head and Neck Squamous Cell Carcinoma Arch Otolaryngol Head Neck Surg. 2009;135(3):261-267. doi:10.1001/archoto.2008.558 Figure Legend: Growth inhibition of head and neck squamous cell carcinoma cells in vitro with 7β-hydroxycholesterol (7β-HC). Cell-Counting Kit 8 (Dojindo Molecular Technologies, Gaithersburg, Maryland) assays were performed on SCC9 (A), SCC25 (B), CAL27 (C), and FaDu (D) cells after 72 hours of treatment with increasing doses of 7β-HC dissolved in dimethyl sulfoxide (DMSO). Cells treated with DMSO served as control. Treatment with 7β-HC resulted in significantly increased rates of cell death compared with control (P < .001). There was near-complete cell death with 25-μmol/L 7β-HC, and 40-μmol/L 7β-HC was required to achieve complete cell death in FaDu cells. Error bars indicate standard errors of the mean. Date of download: 9/30/2017 Copyright © 2009 American Medical Association. All rights reserved.

Copyright © 2009 American Medical Association. All rights reserved. From: 7β-Hydroxycholesterol Induces Apoptosis and Regulates Cyclooxygenase 2 in Head and Neck Squamous Cell Carcinoma Arch Otolaryngol Head Neck Surg. 2009;135(3):261-267. doi:10.1001/archoto.2008.558 Figure Legend: Cyclooxygenase (COX)-1 (left panel) and COX-2 (right panel) expression before and after treatment with 7β-hydroxycholesterol (7β-HC) in SCC9, SCC25, CAL27, and FaDu cell lines. COX expression before treatment is visible in the leftmost lanes (0 hours).Treatment with 7β-HC results in a time-dependent down-regulation of COX-2 in all cell lines (48 hours). A brief up-regulation of COX-2 was detected in SCC25 after 12 hours; COX-1 expression was not influenced by 7β-HC or by tubulin, which served as control. Date of download: 9/30/2017 Copyright © 2009 American Medical Association. All rights reserved.

Copyright © 2009 American Medical Association. All rights reserved. From: 7β-Hydroxycholesterol Induces Apoptosis and Regulates Cyclooxygenase 2 in Head and Neck Squamous Cell Carcinoma Arch Otolaryngol Head Neck Surg. 2009;135(3):261-267. doi:10.1001/archoto.2008.558 Figure Legend: Head and neck squamous cell carcinoma cell lines SCC9, SCC25, CAL27, and FaDu were treated with 10-, 15-, and 20-μmol/L 7β-hydroxycholesterol (7β-HC) for 24, 48, and 72 hours. The cells were labeled with probidium iodide and Anti-Annexin V antibody (BenderMedSystems, Vienna, Austria) and then analyzed with flow cytometry. Graphs show an increase in apoptotic and necrotic cell death in a dose- and time-dependent manner. Error bars indicate standard errors of the mean. Date of download: 9/30/2017 Copyright © 2009 American Medical Association. All rights reserved.

Copyright © 2009 American Medical Association. All rights reserved. From: 7β-Hydroxycholesterol Induces Apoptosis and Regulates Cyclooxygenase 2 in Head and Neck Squamous Cell Carcinoma Arch Otolaryngol Head Neck Surg. 2009;135(3):261-267. doi:10.1001/archoto.2008.558 Figure Legend: Cultured head and neck squamous cell carcinoma (HNSCC) cells. Cultured HNSCC cells (SCC9, A and E; SCC25, B and F; CAL27, C and G; and FaDu, D and H) exposed to the M30 monoclonal antibody after treatment with 15-μmol/L 7β-hydroxycholesterol (E-H). Cells treated with dimethyl sulfoxide served as control (A-D). Cells undergoing apoptosis show cytoplasmatic staining for M30. Clearly, an increase in apoptosis is visible after treatment (original magnification, ×200). Date of download: 9/30/2017 Copyright © 2009 American Medical Association. All rights reserved.