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The single-cell clonal expansion assay was carried out as described (Duan et al., 2015).
Consistent with the observations in growth curve and cell cycle analyses, the results of clonal expansion assay and SA-β-Gal staining also proved early-onset senescence in WS-MSCs (Fig. 4C and 4D).
These cells were grown in vitro and used for in vivo competitive expansion assay.
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Currently available technologies such as fluorescence labeling or clonal expansion assays are almost always end-point analyses of ensemble populations of cells that are expensive, labor intensive, and time delayed.
In vitro expansion assays were performed as described recently [ 23].
Triplet repeat expansion assays using the CAN1reporter were performed as described with the following modifications.
Compared to abi1-3/YFP-ABI1, the ABA sensitivity of the ABAleon2.1 plants was similar in seedling growth assays, but less affected in seed germination and cotyledon expansion assays.
To assess the neuronal differentiation potential along the expansion assays, 2.3 × 106 cells were collected from the suspension cultures and plated in a T75 flask (Nunc).
For in vitro HPC expansion assays, culture-expanded human BM-MSCs (2 × 10 cells/well) were seeded in a 24-well culture plate.
DOI: http://dx.doi.org/10.7554/eLife.01739.011 In seed germination and cotyledon expansion assays ABAleon2.1 lines were hyposensitive to 0.8 µM ABA when compared to Col-0 wild type and YFP-PYR1.
For single cell expansion assays, 24 h after electroporation cells were diluted to a density of 10 cells/ml and plated in 100 μl aliquots into 96 well tissue culture plates.
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