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Forward mapping is performed to replicate the expression profile of BA-BpCAT for verification of endo16 SLM.
Notably, the use of a sense riboprobe corresponding to hybrid grn does not replicate the expression pattern observed for ASgrn1-2.
To replicate the expression in the hermaphrodite and determine expression in the male, we fused the unc-18 upstream sequences used in the mutant-rescue experiment to a YFP reporter.
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This module was validated in an independent material, by replicating the expression changes in allergen-challenged CD4+ cells.
In addition, dsp-4 pretreatment increased inducible nitric oxide synthase expression solely in neurons rather than in microglial cells, more accurately replicating the expression pattern seen in AD patients [ 41].
However, previous reporter analyses showed the POS9 element to be sufficient to replicate the INO expression pattern (but at a lower level of expression) when present in four or more tandem copies [ 12].
The success of the validations depended on the ability to replicate the gene expression detected in the microarray study.
Further, our Transwell experiments reveal that in vitro stimulation of cells does not replicate the gene expression profile seen after in vivo exposure.
Because we were so surprised at not being able to replicate the functional expression of this mutant, we in fact constructed this mutant on three independent occasions, but in each case, the lack of fuctional expression remained the same.
A previous study showed that the POS9 element was able to replicate the endogenous expression pattern only when in combination with additional copies of POS9, or with either the region of P-INO 5′ or the region of P-INO 3′ of POS9 [ 12].
We have recently isolated a genomic DNA fragment ("4.7IL": −3456/+389) that can faithfully replicate the gene's expression pattern.
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