Cell Proliferation Assay with siRNA and Inhibitors
Cells (2 × 104/well) were incubated in 70% MEM with 10% FBS, 30% Opti-MEM with 15 pM siRNA, or miRNA inhibitor (Applied Biosystems) and 0.25 μl/well of Lipofectamine (Invitrogen) in 96-well plates. For the experiments using inhibitors, cells were treated with either SJ001 (cambogin, 5 μM) [65 (link)] or 10058-F4 (an inhibitor that disrupts the c-MYC-Max interaction) [45 (link)–47 (link)] (Calbiochem) at various concentration as indicated in the figures. At 48 h post-treatment, cell proliferation rates were determined using a MTS assay (Promega).
Wang F., Remke M., Bhat K., Wong E.T., Zhou S., Ramaswamy V., Dubuc A., Fonkem E., Salem S., Zhang H., Hsieh T.C., O'Rourke S.T., Wu L., Li D.W., Hawkins C., Kohane I.S., Wu J.M., Wu M., Taylor M.D, & Wu E. (2014). A microRNA-1280/JAG2 network comprises a novel biological target in high-risk medulloblastoma. Oncotarget, 6(5), 2709-2724.
Other organizations :
North Dakota State University, University of Toronto, Hospital for Sick Children, Beth Israel Deaconess Medical Center, Harvard University, Chinese Academy of Medical Sciences & Peking Union Medical College, New York Medical College, University of Florida Health, University of Nebraska Medical Center, Boston Children's Hospital, University of North Dakota
Concentration of inhibitors SJ001 (cambogin, 5 μM) and 10058-F4 (an inhibitor that disrupts the c-MYC-Max interaction)
dependent variables
Cell proliferation rates
control variables
Cell density (2 × 10^4 cells/well)
Culture medium (70% MEM with 10% FBS, 30% Opti-MEM)
SiRNA or miRNA inhibitor (15 pM)
Lipofectamine concentration (0.25 μl/well)
Time point (48 h post-treatment)
Annotations
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