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Dicistronic RNA containing Ex1a, Ex2, or the β-globin 5' leader was transfected into SH-SY5Y cells that were either treated with DMSO (undifferentiated) or retinoic acid (differentiated) for four days (Fig. 9b).
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Nsphs were differentiated for five days in differentiation medium [DMEM/F-12, N2, 1% penicillin/streptomycin and 0.5% FBS (Invitrogen)].
H-2Kb-tsA58 cells were differentiated for five days, to obtain a high yield of differentiated myotubes.
N2a cells were plated and differentiated for two days prior transfection with Httex1-GFP or split-GFP constructs.
Muscle satellite cells were grown on coverslips in 24-well plates, and, for some cultures, differentiated for three to five days into myotubes.
Here we also found that S100β+ astrocytes were present among the cells differentiated for two months from both H9 hESC and TZ1 hiPSC (Figure 4E).
Next, female diploid ES cells were EB differentiated for one or two days, and then subjected to γ-irradiation-, mimosine- or colcemid-mediated cell cycle arrests for one day.
ES cells were EB differentiated for one or two days and then blocked in the cell cycle by adding 0.75 mM mimosine, 12 µl/ml colcemid (KaryoMax, Gibco) or 2100centiGray γ-irradiation.
To exclude the possibility that our triploid XXY ES cells lost or gained X chromosomes over the differentiation process, we performed DNA-FISH with an X chromosome specific BAC probe on cells differentiated for seven days.
In striking contrast to the mC2C12 cells, C2C12 cells that had been differentiated for nine days (dC2C12) did not undergo death when injected with bovine or yeast cytochrome c (Fig. 1a, b).
The third method of assessment was performed with PC12 cells being differentiated for five days in the presence of NGF and then exposed to viral vectors for three days in the continuous presence of NGF, followed by withdrawal of NGF and serum.
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