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Differences in the median reporter values between the wildtype and mutant variants of PPP1R15A were evaluated by One-way ANOVA, by the medians Kruskal Wallis test (cells not expressing mCherry and with an intensity above 10 were discarded) using GraphPad Prism version 6.0e (GraphPad Software).
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In all luciferase experiments cells were co-transfected with a renilla luciferase control vector (pRLTK, Promega) and reporter firefly values were normalised to renilla values.
Note that there is no necessary correlation between the reporter p-value and p-value of the expression change in the level of the regulator itself.
For the synthetic reporter, the values were scaled to that of the 3X-AP1 site reporter.
Overexpression of miR-4732-3p in K562s resulted in a significant reduction in relative reporter luciferase values (Fig. 5a), demonstrating that miR-4732-3p suppresses SMAD4-mediated transcription activitranscription
A graphical representation of the results obtained with RootReporter can be observed in Figure 4 figure supplement 1. Reporter intensity values along the first 5 mm of root tips can also be observed in Figure 4 figure supplement 2. 10.7554/eLiFigure97.025 Figure 4. Dual-color reporter visualization of root structure and gene expression.
All the features presented in heatmaps are those features that have reporter P-value < 0.001 and P-value < 0.01 were considered for INO-level and deficient factors, respectively.
Reporter p-value indicates the significance of the average change in the regulated genes.
The highest Rn (normalized reporter fluorescence) value with the lowest primer concentrations is the indicator by which one selects the appropriate optimal primer concentrations in each case.
In all luciferase experiments, transfection efficiency and cell viability were controlled for by cotransfecting a Renilla luciferase control vector (pRLTK; Promega, Southampton, UK), and reporter firefly luciferase values were normalized to the Renilla values.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com