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Lastly, the BIN1-Tau PLA staining partly co-localized (mean ± standard deviation (SD) co-localization coefficient: 0.399 ± 0.041) with the actin cytoskeleton (as revealed by phalloidin staining; Fig. 3e, f, Additional file 10 and Additional file 11).
Likewise, specific pre- or post-synaptic markers of functional neuronal compartments (such as synaptophysin (Fig. 4b) and PSD95 (Fig. 4c)) did not co-localize with BIN1-Tau complexes (mean co-localization coefficient ~0; see Additional file 10).
The weighted co-localization coefficient represents the sum of intensity of co-localizing pixels in channels 1 and 2 as compared to the overall sum of pixel intensities above threshold.
The co-localization coefficient of 0.87 between Mito-MQ and MitoTracker Red revealed that the probe was predominantly present in mitochondria, therefore, Mito-MQ was successfully applied to detect mitochondrial oxidative stress by detecting the change of Cys/Hcy.
Meanwhile, standard co-staining experiments of MPCB and Mito Tracker Red FM (co-localization coefficient: 0.89) revealed that MPCB can be used for the real-time sensing and bioimaging of SO2 derivatives in mitochondria.
The co-localization coefficient represents the weighted co-localization coefficients of Ch1 (red) with respect to Ch2 (green) for each experiment [57].
Given that the co-localization of the two proteins increased substantially in fibers treated with the proteasome inhibitor, as determined by the co-localization coefficient (Fig. 4B and D), we can infer that Ozz regulates the proteasomal degradation of a selected pool of sarcomeric MyHCemb during myofiber differentiation.
The co-localization coefficient was calculated according to Mander's method using ZEN 2012 software.
Co-localization was measured by Pearson Co-localization Coefficient using Volocity Image analysis software.
This was reflected in the lower co-localization coefficient of the Lifeact and SEPT2-GFP probes in NFs.
A threshold value of 40 was used and the Pearson's co-localization coefficient was recorded for each neuron analysed.
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