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GUS staining was performed on stable transformants as previously described [12].
VIGS experiments used the purple-flowered 'Primetime Blue' cultivar, but studies on stable transformants used white-flowered cultivar 'Mitchell Diploid'.
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To select for stable transformants, transformations were cycled on and off antibiotic selection for three 1-week intervals.
On the other hand, 20 87% of stable transformants were targeted using an AAV-NANOG-targeting vector designed for the promoter-trap strategy.
Both of these constructs also harbored a neomycin-resistance marker to facilitate isolation of stable transformants.
Selection of stable transformants.
For the generation of stable transformants, the infected cells were selected with 1 μg/mL puromycin.
To select for stable transformants, regenerating protoplasts were cycled on and off antibiotic plates for three 1-week intervals.
In addition, we used qRT-PCR on some of the stable transformants expressing STOX1 at different levels, and obtained an external validation of the microarray results (Figure 2).
Stable transformants were selected on YPD medium containing nourseothricin.
Stable transformants were selected on 25 μg mL−1 hygromycin.
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