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Average mean puncta intensities were calculated for each animal, and then for the population of animals.
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Representative images (C, scale bar 5 μm) and mean puncta intensity (D ) are shown.
Representative images (B, scale bar 5 μm) and mean puncta intensity (C ) are shown.
(E ) GFP-UNC-49B synaptic abundance was unaltered in nrx-1 mutants Representative images (top, scale bar 5 μm) and mean puncta intensity (below) are shown.
The shift in GLR-1 localization from small puncta to large accretions can be quantified by measuring mean puncta size in the ventral cord (Fig. 1I).
(B and C ) Cumulative probability plots of VGAT and PV puncta intensities in control and Aph1bc −/− null mice.
(G and H ) Cumulative probability of VGluT1 and Homer1 puncta intensities in control and Aph1bc −/− mice.
Moreover, the cumulative distribution of surface GluA2 puncta intensities was right-shifted in Vac14 −/− neurons.
The charts in Figure 3B show analysis of a minimum of 25 cells per condition and count puncta per cell, and mean puncta diameter.
Finally, to confirm our results were not an artifact due to how the autophagosome score was defined (ratio between puncta intensity and the total cellular GFP-LC3 intensity), we also tested the distributions of other ways of scoring for autophagy, including number of puncta, total area of puncta and average brightness of puncta.
In order to quantitatively confirm the results, we used image analysis software to determine the mean intensity of puncta at each compound concentration.
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