Fig. 1.

C2C12 cell adhesion after NaBC1 and fibronectin-binding activation. a) Immunofluorescence images of actin cytoskeleton (green), nuclei (blue) and vinculin (magenta) as a marker of focal adhesions. C2C12 cells were cultured for 0.5, 1 and 1.5 h onto fibronectin-coated glass substrates (Glass), and in the presence of 0.59 mM borax (Glass-B). NaBC1 activation induced clear focal adhesions as a result of vinculin staining at the different time points. Scale bar 50 μm. b) Image analysis quantification of different parameters related to cell shape, spreading area, focal adhesions (FA) and total actin stress fibers. NaBC1 activation induced more focal adhesions number and area after 0.5, 1 and 1.5 h. Bigger focal adhesions are present after NaBC1 activation after 0.5 h of culture. Active-NaBC1 induces actin stress fibers formation at the different time points evaluated. Statistics are shown as mean ± standard deviation. n = 15 images/condition from three different biological replicas. Data were analyzed by an ordinary two-way ANOVA test and corrected for multiple comparisons using Tukey's correction analysis (P = 0.05). In the cell parameters graphs, statistics indicate differences among the different time points evaluated (there is no difference between columns at the same time point). In the focal adhesions parameters and actin graphs, statistics indicate differences between columns of the same condition. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.) (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)