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A second portion of the collected cells was used for plating on glycerol-based and glucose-based solid medium lacking drug to assess the functionality of mtDNA.
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In the initial screening we identified 218 colonies that grew very slowly, or not at all, on medium containing antimycin A while exhibiting obvious growth on medium lacking the drug.
Cells incubated for 72 h with complete medium lacking antiviral drugs were included as viable controls.
Medium lacking serum was termed 'basal medium' (BM), while medium containing 10% FBS was termed as "full medium" (FM).
The resistant phenotype was not stable if cells were transferred to complete drug-free medium, but remained stable for at least 3 months in the presence of medium lacking oestrogenic activity.
The medium was exchanged with fresh medium lacking phenol red.
A functional Ura3 protein is produced, enabling selection on medium lacking uracil for the parental vector.
Transformants were selected on medium lacking leucine at 32 °C to obtain single colonies.
For negative controls, unstimulated neutrophils in regular RPMI medium lacking phenol red were used.
In chase experiments, cells were pulsed for 30 min with radiolabel then the medium removed and replaced with fresh medium lacking the radioisotope for the indicated time.
Leu+ transformants were selected on SC medium (synthetic complete medium) lacking leucine (SC-Leu).
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