Pll coated 96 well plates

Cells were seeded onto poly-l-lysine (PLL)-coated 96-well plates (Greiner Bio-One, Kremsmünste, Austria) at 8000-10,000 cells/well with Micro Shot 706 (MSTechnos, Tokyo, Japan). They were cultured in DMEM supplemented with 10% FBS and 300-600 µM mexiletine for 48-72 h. The quantity of the culture medium was unified with 100 μL. A calculated volume of baculoviral vector solution was directly added to cells in the wells approximately 24 h before the assay. On the assay, the cells were washed with the bathing solution using Multi Works 508 (MSTechnos) in the following manner. Two hundred microliters of the bathing solution was slowly poured into the wells, and then 200 μL of the solution was removed. This procedure was repeated six times for washout of mexiletine. After the washout, 100 μL of bathing solution that contains test compounds and BaCl 2 was added to each well to bring the final concentration of Ba 2+ to 200 μM. Cells in 96 wells were incubated for 12-16 h at 37 °C, and then proceeded to MTT assay. Plates were analyzed for absorption on ARVO MX (1420 Multilabel Counter; PerkinElmer, Waltham, MA).