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Third instar larvae were irradiated and cultured on medium containing drug.
The transfection medium was removed and fresh culture medium, containing drug or carrier control, was added.
Medium containing drug was aspirated and fresh growth medium containing 0.5 mg/mL MTT was added to cells.
The next day, medium was replaced by fresh medium containing drug as indicated or DMSO, each condition in triplicate.
Twenty-four hours later, cells were treated with drugs at the specified doses and incubated in normal medium containing drug for 12 days, after which they were fixed in 90% methanol, and stained with 0.1% crystal violet.
PSMB7 overexpressed in resistant cells was silenced in doxorubicin-resistant MCF-7 cell lines (MCF-7-RAdr); 24 h after transfection, the reaction was stopped by replacing the medium with normal growth medium containing drug in concentration of 0.2 μg ml 1.
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Medium was replaced with medium containing drugs at varying concentrations, and the cells were then incubated at 37 °C for 24 h before 10 μl of CCK-8 (Dojindo Laboratories, Rockville, MA, USA) was added to each well.
After repeated several cycles and when the cells grew up to 70% confluence in the culture medium containing drugs, the concentration of drugs will increased to the next level.
The cells were diluted to an OD600 of 0.2, and 4 µL was spotted with serial dilutions on solid medium containing drugs (AmB, fluconazole, itraconazole or ketoconazole) in the agar.
Drug treatments were accomplished by aspirating the medium and replacing it with new medium containing drugs.
For the long-term treatment of cells, NSC or complete medium containing drugs was replaced every 2 3 days.
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medium containing galactose
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medium containing dye
medium containing chemical
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medium containing control
medium lacking drug
medium containing chloramphenicol
medium containing serum
medium containing antibiotic
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