Rpmi 1640 dmem media l glutamine

PC-3 and WISH cell lines were obtained from the National Cancer Institute in Cairo, Egypt, cultured in “RPMI-1640/DMEM media l-Glutamine (Lonza Verviers SPRL, Belgium, cat#12-604F). The cells were cultured in 10% fetal bovine serum (FBS, Sigma-Aldrich, MO, USA) and 1% penicillin-streptomycin (Lonza, Belgium)”. All cells were incubated at “37 °C in 5% carbon dioxide atmosphere (NuAire)” following routine tissue culture work. Cells were seeded in triplicate at a density of 5 × 10⁴ cells per well in a 96-well plate, and then treated with the compounds at concentrations of 0.1, 1, 10, and 100 μM the next day. Cell viability was assessed using MTT solution (Promega, USA).^{60 (link)} The plate was incubated for three hours. The absorbance was then measured with an ELISA microplate reader (BIO-RAD, model iMark, Japan). GraphPad Prism 7 was used to determine IC₅₀ values based on survival rates relative to a control group, as previously reported in ref. 61 (link).