Variability in dystrophin quantification in independent immunostaining and dystrophin signal detection stability across the time.

<p>Double staining with different antibody combinations were performed (anti-dystrophin ab15277/anti spectrin (a) and anti-dystrophin MANDYS106/anti-laminin (b)). Sections were cut and stained immediately (A). Muscle blocks were kept at -80°C for one month and then two section sets were cut and again stained immediately. One set of these stained sections were acquired immediately (B) whereas the other stained section set was kept at 4°C for three months and then acquired (C) in order to evaluate the fluorescent signal stability of dystrophin staining after slides long time storage. Dystrophin intensities were quantified per each individual fibre and were plotted as scatter plots (mean±SEM) in arbitrary units (***p< 0.001, Mann Whitney test). Dystrophin intensity dynamic range and number of fibres acquired per experiment are reported in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0194540#pone.0194540.t003" target="_blank">Table 3</a>.