Fig. S3

Damage response of vesicle markers. (A) Lysosome-associated protein transmembrane 4a (Laptm4a) or lysosome-associated membrane protein 2 (Lamp2), fused to mTFP1 in their C-terminus (green) were co-expressed with prenylated mCherry (magenta). Cells were damaged (arrow) with the two photon laser and localization of lysosomal markers was recorded. No large-scale redistribution of lysosomes was evident, although limited amount of mTFP1 could occasionally be observed near the lesion. (B-C) Quantification of lysosome dynamics at the site of lesion. Fluorescence change (%) relative to 20 s time point after injury is depicted. (D) Lysosomes (arrowhead) were observed near the lesion (arrow) only when the rupture occurred close to them. Even then, they did not relocate and fuse to the membrane. (E) Quantification of various endocytosis or exocytosis markers at the site of sarcolemmal lesion. The vertical axis demonstrates fluorescence change at the site of lesion (%) relative to the 20 s time point. Due to the irregular distribution of the vesicles across the cell, the analysis was carried out relative to the 20 s time point. By this time mTFP1-DysfC had already significantly increased at the site of lesion, yet still continued to accumulate also afterwards. Such clear trend could not be observed for any of the studied vesicle markers. Scale bars: A, 4 μm; D, 2 μm