Figure 5.

Annexin A2 distribution in dysferlinopathy myoblasts. Distribution of annexin A2 was analyzed by immunofluorescence using an anti-annexin A2 antibody and Cy3-conjugated anti-rabbit secondary antibody in C25 and DYSF3 myoblasts permeabilized with 20 µM digitonin in the presence of 300 nM Alexa-Fluor-488 actin, 2 mM ATP-Mg²⁺, and 10 µM free Ca²⁺. Confocal images were captured at the equatorial plane of the cells using identical exposure settings between compared samples; therefore, annexin A2 distribution was measured in a single focal plane. (a) C25 and DYSF3 myoblasts immunostained with annexin A2 (red) and fluorescent G-actin incorporated into filaments (green). Scale bar = 20 µm. Insets show digital magnification of the boxed areas. (b,c) Bars represent means ± SEM of the ratio of the mean fluorescence intensity of the cytosol/whole cell of annexin A2 immunostaining (b) and Pearson correlation coefficient for colocalization of annexin A2 with fluorescent actin filaments (c). The number of analyzed cells from four different cultures is indicated in parentheses. * p < 0.05 (t-test).