Figure 4.

Recruitment of AnxA6 to the damaged membrane site and formation of the cap subdomain in LHCN myotube. (A) LHCN myotubes were transfected with the plasmid pA6-GFP and membrane damage experiments were performed by laser ablation at the lateral edge of the myotube. Red arrow, area before irradiation; white arrow, area after irradiation. Scale bar: 10 µm; (B) Magnified images of the disruption site before (left) and more than 65 s (right) after laser ablation. The dashed white line corresponds to the position of the plasma membrane before irradiation. Scale bar: 5 µm; (C) Distribution of endogenous AnxA6 in a damaged LHCN myotube. The plasma membrane of LHCN myotubes was damaged by laser irradiation. Myotubes were immunostained for AnxA6 and observed in bright-field (left) and fluorescence microscopy (right). White arrow, irradiated area. Scale bar: 5 µm; (D) Distribution of AnxA6 and dysferlin between cap and shoulder subdomains. A LHCN myotube expressing AnxA6-GFP (green) was damaged at the lateral edge and then fixed and immunostained for dysferlin (magenta). Red arrow, irradiated area (E) LHCN myotubes were transfected with the plasmid pA6-GFP and membrane damage experiments were performed by laser ablation on the surface of the myotube. Red arrow, area before irradiation; white arrow, area after irradiation. Scale bar: 20 µm.