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ClC-3, which could be detected in western blot analysis (Fig. 2D) (possibly because of endosomal contamination of the membrane fractions) could together with endogenous ClC-7 be responsible for the small endogenous background currents in the CHO control preparations (Fig. 3A).
High extracellular K+ was used to shift the resting membrane potential toward 0 mV and avoid voltage dependent changes in TRP activity due to off target effects on endogenous background currents (the SNAP-induced conductances observed in whole cell experiments with HEK293T cells reversed at +15±5 mV, n = 3).
Termination of the kinase reaction at a certain timepoint revealed recombinant kinase activity (wt AND opt) clearly over endogenous background activity (mock), while higher amounts of "optimized" protein resulted in higher amounts of phosphorylated substrate (Fig. 4C, lower blot and panel).
Epididymis was excluded because of endogenous background staining (Fig. S4I).
The incidence, location and range of intensity of endogenous background was extensively assessed in wild-type mice (n=116).
These reports support the role of MMP-1 promoter polymorphism as an endogenous background factor, in contrast to exogenous environmental factors [ 15].
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Thus we elected to transfect LNCaP cells with different Abi1 isoforms and to compare the function of these isoforms against a very low endogenous Abi1 background.
The dominant experimental paradigm in functional magnetic resonance imaging (fMRI) assumes that the brain's response to an experimentally controlled task is independent of its endogenous or background oscillatory activity.
Negative control PLAs (GFP AMPKα1) detecting non-expressed GFP and endogenous AMPKα1 background are also displayed.
The percentage of quench-protected toxin fluorescence above the endogenous cellular background level (BG) is indicated.
In either case, these results indicate that materials other than chlorophylls are responsible for endogenous green background fluorescence.
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