Fig. 5

Silencing of dysferlin induces the presence of connexin 40.1, 43 and 45 in human myotubes. Control RCMH myoblasts were transfected with small hairpin RNA to decrease dysferlin expression, to be later differentiated to myotubes by culturing in differentiation medium for 14 days. Once myotubes were obtained, the presence of connexin 40.1, 43, 45 (green signal) and dysferlin (red signal) was analyzed by immunofluorescence assay using specific antibodies and compared with that of control RCMH myotubes (without transfection). The three connexins, but not dysferlin, were detected in RCMH-sh Dysferlin myotubes, whereas control RCMH myotubes present dysferlin (red signal) but not Cxs. The nuclei were stained with DAPI (blue signal). Scale bar: 50 μm. n = 4 cell cultures for each cell line