RPMI 1640, 1X

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	Supplemented with 10% fetal bovine serum

Protocol tips
Supplemented with 10% fetal bovine serum

Publication protocol

DU145 and LNCaP cells were maintained at low passages (below 30) and cultured in RPMI 1640 (Wisent Bioproducts, QC) with 5 and 10% fetal bovine serum, respectively. For MTT proliferation assays 3500 LNCaP (on poly-L-lysine coated wells) or 1500 DU145 cells were seeded in 96-well plates, and after 24 h, the cells were treated with various concentrations of inhibitors (from 300 to 1 μM). Next, the cells were incubated for 72 h with peptides prior to addition of MTT reagent (Sigma–Aldrich, Canada) at a final concentration of 1 mg/mL. Formazan salt was solubilized with 50 μL DMSO and the metabolic activity was normalized relatively to vehicle-treated cells (sterile bi-distilled water). IC50 values were determined using Prism 5.0 (GraphPad Software, USA), as previously described12. For LDH assay, the LDH Cytotoxicity Detection Kit (Promega, USA) was used to quantify the release of lactate dehydrogenase following membrane disruption. As described in our previous work15, DU145 cells were seeded at a density of 3000 cells/well and allowed to incubate for 24 h. Then, the media was subsequently replaced and cells were incubated with compounds at the selected concentration. Cells treated with 1% SDS were used as a control for maximal LDH release and further relative quantification.

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