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Blood, Vol. 112, Issue 3, 760-769, August 1, 2008
Cyproheptadine displays preclinical activity in myeloma and leukemia
Blood Mao et al.
112: 760
Supplemental materials for: Mao et al
Files in this Data Supplement:
- Figure S1. Cyproheptadine decrease cyclin D2 transactivation through a c-maf independent mechanism (JPG, 13.6 KB)
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NIH3T3 cells over-expressing c-maf and the cyclin D2 promoter-driving luciferase (squares) or NIH3T3 cells over-expressing the cyclin D2 promoter-driving luciferase without detectable levels of c-maf (diamonds) were plated in 96-well plates (13,000 cells per well). After the cells had adhered to the plates, they were treated with increasing concentrations of cyproheptadine. Cells were incubated with increasing concentrations of cyproheptadine for 20 hours. After incubation, cyclin D2 transactivation was assessed by the luciferase assay as described in the Materials and Methods. In parallel, we also assessed viability with an MTS assay. The relative activity represents mean (cyproheptadine luciferase RFU/control luciferase RFU)/(cyproheptadine MTS OD/control MTS OD) ± SD.
- Figure S2. Cyproheptadine induces cell death by Annexin V staining (JPG, 18.6 KB)
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KMS11 and LP1 myeloma cells were treated with 20 µM of cyproheptadine. At increasing times after incubation, cells were harvested and the percentage of Annexin V positive cells determined by flow cytometry. Data represent the mean ± SD percent of Annexin V positive cells.
- Figure S3. Cyproheptadine induces cell death independent of its inhibition of the histamine and serotonin receptors (JPG, 49.4 KB)
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OCI-AML2 and CEM leukemia, OPM1 and KMS11 multiple myeloma cells were pre-treated with 10µM of histamine (H), serotonin (S), or histamine and serotonin, or buffer for 24 hours. After pretreatment, cells were incubated with increasing concentrations of cyproheptadine (CYP) for an additional 48hr. After incubation, cell viability was measured by MTS assay. Cell viability is expressed as a mean percentage ± SD relative to untreated cells.
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