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The background fluorescence level at each concentration was determined by setting up serial dilutions of 0 370 μM Rpn10 in HBS.
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Mice inoculated with TCR-NYE-wt exhibited only transient tumoral fluorescence at the 4- and 8-h imaging time points before a return to background fluorescence levels at 48 h (Fig. 3b, c).
The gray region is the background fluorescence level used to define the significance threshold (mean ± 2.5 s.d.).
The background fluorescence level for each specimen was established using the isotype control.
Both groups shared a certain background fluorescence level after OGD or TBI.
Background fluorescence levels were determined by Fluorescence Minus One (FMO).
After 100 s of destaining, a second stronger stimulation (2000 stimuli at 20 Hz without imaging (for 100 s)) was carried out to reach background fluorescence levels.
In order to determine the area of individual growth cones, we set fluorescence thresholds slightly above the background fluorescence levels and generated binary images (background fluorescence = 0, membrane fluorescence = 1).
As TRPV1 channels opened, the background fluorescence levels gradually increased during the course of the video.
For this analysis, exposures were adjusted for background fluorescence levels using untransfected cells.
Only with mice injected with Alexa 680 labelled high-affinity TCR-NYE- 0.048 TCR-NYE- 0.048 TCR-NYE- 0.048nM fluorescence levels observed throughout the time course; significantly, higher fluorescence intensities were noted in thigherice athan and 48 h (p < 0.05), and even at 96 h, intensities were 2.5 times higher than background.
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