Verapamil hydrochloride v4629

Metabolic activity of cells as a marker of cell viability and proliferation was assessed by WST-1 Cell Proliferation Assay Kit (10,008,883, Cayman Chemical, MICH, USA) according to the manufacture’s recommendation. Three MB cell lines, D341, CHLA-01 and CHLA-01R, were used in this assay. Briefly, cells were seeded at a density of 30,000 cells/well in 96-well plates. Twenty-four hours later, treatment was added as shown in the “Results” section. Assay reaction was assessed 24 or 72 h after treatment. For that, 10 μL of WST-1 reagent was added to each well for 4 h, and absorption was measured using a plate reader (Multiskan Go, Thermo Fisher Scientific, Scoresby, VIC, Australia). Drugs used, including mibefradil dihydrochloride hydrate (M5441), NNC55-0396 hydrate (N0287), verapamil hydrochloride (V4629), nifedipine (N7634), vincristine (V0400000) and lomustine (L5918), were purchased from Sigma-Aldrich.

Doxorubicin (D4035), etoposide (E1383), cisplatin (P4394), verapamil hydrochloride (V4629) and cyclosporin A (C3662) were obtained from Sigma-Aldrich.