A proliferation assay was performed using the colorimetric CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) (Promega) on PC-3 and A-431 according to instructions from the manufacturer (DSMZ, Braunscweig, Germany). In brief, the two cell lines were treated with 2, 4, 8 and 20 μM of Perhexiline Malate salt (Sigma-Aldrich, St. Louis, USA) for 24 and 48 hours respectively. Perhexiline was dissolved in DMSO, and corresponding concentrations of DMSO in medium were used as controls. In addition, controls consisting of cells growing in only cell culture medium were included. For all concentrations and corresponding DMSO-controls eight replicates were analysed, and for the controls consisting of cells in only medium 16 replicates were analysed. In the follow up experiment the two cell lines were treated with 4 and 6 μM respectively of Perhexiline Malate salt (Sigma-Aldrich, St. Louis, USA) for 48 hours. This time 16 replicates were used for all conditions and controls.
Ghaffari P., Mardinoglu A., Asplund A., Shoaie S., Kampf C., Uhlen M, & Nielsen J. (2015). Identifying anti-growth factors for human cancer cell lines through genome-scale metabolic modeling. Scientific Reports, 5, 8183.
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