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This assay has the advantages that it is simple in design, produces robust behaviors, and enables the collection of data from large numbers of animals in a short experimental timeframe.
In our simulations (Fig. 3), we assume that the system reaches a steady state within the experimental timeframe of 2 hours.
Prior to wider pharmacological validation of this approach, the variability of each quantifiable parameter was assessed over the experimental timeframe.
A total flux was calculated as area between flux curve over the indicated experimental timeframe (25 min) and the starting flux value.
Within the experimental timeframe, almost all induced genes were induced by 4 h post-exposure.
Furthermore, relative levels of bI4 maturase protein remained at least two-times higher during the entire experimental timeframe.
The rapid experimental timeframe of iNeuron generation and the potential to reprogram to specific neuronal subtypes make this an appealing experimental strategy for in vitro models of neurological disease.
This finding indicates either a long intracellular half-life for SMAD1 in USSC, unaltered by miRNA transfection in the experimental timeframe, or secondary regulatory mechanisms such as increased transcription that keep SMAD1 levels constant.
Advantages of the CER assay include its low cost, short experimental timeframe and replacement of living animals with tissue samples (which need not be from transgenic mice or perfused).
Dextran flux was assessed to determine the experimental timeframe of 6 hours, in agreement with previous work with large molecule transport [ 26]. 6 hours prior to the experiment, cell medium was changed to 2% FBS. 10 μg/mL adiponectin and dextran were applied above the monolayer (luminal side) to the top well.
A: Experimental.
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