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Nonlinear regression (sigmoidal dose response) was used to calculate the EC50 values and maximal fold activation.
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Up to 3 fold activation was observed in HEK293T cells, with maximal activity when lower amounts of CRTR-1 plasmid were used (Figure 1B).
It activated the receptor with an EC50 value of 3.9 μM and reached maximally two-fold activation compared to 12-fold maximal activation with an EC50 value of 0.3 μM by the full agonist pioglitazone in the same system (Fig. 2B).
The results were presented as fold activation over the control.
We determined that caspase 8 activity increases in a time-dependent manner with a 2-fold maximal activation after roughly two hours of culture in DM (Fig. 1(A)).
The number of genes with large maximal fold-change between mice (D).
The kinetics of β-galactosidase levels shows that an ∼10-fold activation of gene expression was reached after ∼4 h, and the maximal ∼40-fold activation was reached after ∼8.5 h of irradiation.
Maximal activation of AMPK in WT cells was observed at 10 min after which it declined, although >2-fold activation was still observed at 60 min (Fig. 2C).
While yeast-secreted Ex4 showed the highest activation by 59-fold, GLP1 and S2-GLP1 secreted by yeasts provided only 5.8 and 1.7-fold activation, respectively.
The results showed that 4 × 104 yeast cells were enough to detect GLP1R activation, showing 73-fold activation, compared with the control, and 1 × 106 yeast cells provided the largest dynamic range with 224-fold activation of GLP1R.
Osmoresponsive transcription is characterized by strong induction of gene expression from almost no expression under basal conditions to maximal activation (fold changes ranging from 5 to 200) in just 10 min (Posas et al. 2000).
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