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Samples were prepared in 96-well plates containing 0.06 ml per well of respiratory buffer.
Alternatively, in a 24-deep-well plate, LB can be used (6 ml per well) for overnight cultures to increase the final DNA content.
Culture medium was replaced with 2 ml per well of culture medium containing liposomal solutions (30 μg DOX/ml).
The plate was incubated at 37°C for 72 hrs in 5% CO2 incubator with the final volume of 0.2 mL per well.
To generate DCs, the mononuclear cell fraction was washed twice with RPMI-1640, suspended in RPMI-1640 at 2.5 × 106 cells/mL, and seeded in a 6-well plate (Becton) at 2 mL per well.
Cells were expanded in 24-well plates (105 cells per well) in culture media (0.5 ml per well).
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In parallel, 2E6 FLuc-expressing SKOV3 cells were imaged upon adding 150 μl of d-luciferin (30 mg/ml) per well.
In each assay, the Escherichia coli fosmid clones, pre-grown overnight (96-well plates, 200 μl LB + chloramphenicol (12.5 μg/ml) per well, 37 °C, 200 rpm), were inoculated onto the respective agar medium (with 0.01% arabinose).
After drug treatment for 72 hrs, 50 μL MTT solution (2 mg/mL) per well was added and incubated at 37°C.
The cultured medium was removed and replaced with 150 μl MTT (0.5 mg/ml) per well before termination at 4 h.
HT29 cells were seeded into 96 well tissue culture plates at a density of 2×105 cells/mL per well.
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