Moreover, an increase in reporter activity was also observed with increasing amounts of transfected receptor (50 ng to 500 ng).
Consequently, in response to agonist stimulation of G-protein-coupled receptors (GPCRs), the engineered yeast achieved more than a 150-fold increase in reporter intensity in up to 98% of cells, as determined by flow cytometric sorting.
Treatment of the orthotopic xenograft with varying doses of erlotinib (25, 50, and 100 mg/kg) revealed a dose- and time-dependent increase in reporter activity (10-, 12-, and 23-fold).
We found that erlotinib treatment of resistant A549 subcutaneous and orthotopic xenografts resulted in significant AKT inhibition as determined by an 8- to 13-fold (P <.0001) increase in reporter activity 3 hours after erlotinib (100 mg/kg) administration compared to the control.
Transverse aortic constriction of adult mice harboring ANF reporter transgenes demonstrated no significant increase in reporter activity in the ventricle.
As expected, the luciferase activity was markedly reduced in the cells transfected with pre-miR-10a, whereas the inhibition of miR-10a resulted in an increase in reporter activity compared with transfection with anti-miR control (Fig. 3E).
While microRNA cocktail reprogramming on traditional 2D tissue culture plates produces a five-fold increase in αMHC-CFP reporter expression, reprogramming in 3D fibrin hydrogels generates a twenty-fold increase in reporter expression (Li et al., 2016).
As expected, overexpression of miR-495 resulted in approximately a 20% reduction in luciferase reporter activity, whereas inhibition of miR-495 resulted in a 1.3-fold increase in reporter activity compared with the cells transfected with control inhibitor (Fig. 2C).
As expected, miR-19a/b overexpression resulted in a ~40% reduction in luciferase reporter activity compared to cells treated with the pre-miR control, whereas miR-19a/b inhibition resulted in a 1.5-fold increase in reporter activity compared to cells transfected with the anti-miR control (Fig. 3E).
As expected, miR-181b overexpression resulted in an approximately 70% reduction in luciferase reporter activity compared to cells transfected with control mimic, whereas miR-181b inhibition resulted in a 30% increase in reporter activity compared to cells transfected with the control inhibitor (Fig. 3E).
miR-124, however resulted in significant increase in reporter gene expression.
