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Fig. 5
The WRRFK-motif is required for accumulation of Dysf and PS.
(a) After membrane damage, zfWRRFK-TM-C (red) but not zfWAAFK-TM-C (grey) accumulated rapidly at the lesion. The effect was reduced on exchange of the first (zfWARFK-TM-C; purple) or second arginine by alanine (zfWRAFK-TM-C; green). (b) PS (LactC2:GFP) enrichment in the repair patch of Dysf-KD myofibers co-expressing zfWRRFK-TM-C (green), but not of those co-expressing zfWAAFK-TM-C (red). (c-e) HeLa cells expressing zfWRRFK-TM-C were imaged before (c) and 0 s (c, arrow) and 270 s after membrane damage (c, arrows). zfWRRFK-TM-C markedly amassed at the lesion (c, arrow, inset; e, black), whereas zfWAAFK-TM-C showed only baseline fluorescence recovery after photobleaching (d, arrows, inset; e, red). (f-h) HeLa cells transfected with LactC2:GFP and imaged before (f) and 0 s (arrow) and 270 s after lesioning (arrow) showed no PS accumulation but only baseline recovery (f, arrow, inset; h, red); in the presence of zfWRRFK-TM-C, LactC2:GFP accumulated within the repair patch (g, arrows, inset; h, black). The data in (a,b,e,h) are given as mean±s.e.m. (n=>5) and scaled such that 100% corresponds to fluorescence from the same area before damaging. Scale bars, 10 µm (c,d) and 12.86 µm (f,g).