Cells were passaged the day before electroporation. We washed about 5 to 10 million cells with PBS twice. Opti-MEM medium (500 μl) was added to the resuspended cells. To deliver a single DNA plasmid, we added 15 μg of plasmids to the resuspended cells. To introduce two or three plasmids, we added 10 μg of DNA for each plasmid to the resuspended cells. We then transferred the mixture to the 4-mm cuvette (Bio-Rad) for electroporation (270 V, 950 μF, infinite resistance) while avoiding bubbles. After electroporation, cells were transferred from the cuvette to the 35-mm dish with 5 ml of transfection medium for culture in the incubator.

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