Fig. 3
Five amino acid motif in Dysf is required for PS accumulation. (a,b) Before wounding, LactC2:GFP (green) localized to the sarcolemma (sc; a) and the Z-line (z; a) both in control (a) and knock-down (dysf-KD; b) myofibers. On injury, LactC2:GFP accumulated in controls (a, arrow) but not in dysf-KD myofibers (b, arrow). (c) Kinetics of LactC2:GFP accumulation in control-KD (green), dysf-KD (red) and dysf-KD embryos co-injected with mOrange1-DysfC (Dysf-mO, yellow). Lack of PS accumulation in dysf-KD myofibers was rescued by Dysf-mO, translation of which is not inhibited by dysf morpholino. (d) Knock-down of anxa6 (anxa6-KD) had no effect on PS accumulation (red) compared with controls (anxa6-control, green). (e) Domain structure of Dysf. (blue AA: predicted amphipathic helix). (f,g), On damage zfWRRFK-TM-C accumulates in the repair patch (f, arrow). No accumulation was observed for zfTM-C (g, arrow). (h) Accumulation kinetics of zfWRRFK-TM-C (red), zfWRRFK-TM (green) and zfTM-C (blue). A 5-AA motif (WRRFK, red box, e) is required for accumulation. (i) zfWRRFK-TM-C (red) but not zfTM-C (green) rescued PS accumulation in dysf-KD myofibers. In all charts, the change of fluorescence at the lesion is indicated as percentage relative to the undamaged state±s.d.). Scale bars, 4 µm (a,g-h), 3 µm (b). |