Fig. 4. Immunostaining of WT and heterozygous targeted cells shows normal localisation of obscurin and telethonin, but some mislocalisation of MURF2. WT and heterozygous cells were coimmunostained for obscurin and -actinin, Z-disk titin (T12 antibody) and telethonin, or for Z-disk titin (T12 antibody) and MURF2 and imaged using a confocal microscope. The arrows show the alternate localisation of obscurin at the M-band and -actinin at the Z-disk in the top pair of panels for WT and heterozygous cells (clone 5-A8). Titin and telethonin are both localised to the Z-disk (arrows in the middle panels) for WT and heterozygous cells (clone 1-F9). There is also a diffuse cytoplasmic pool of telethonin. In the bottom pair of panels, the arrows show the alternate localisation of titin at the Z-disk and MURF2 at the M-band in WT cells, but this alternate localisation is less clear in the heterozygous cells (clone 5-A8). Arrows indicate the M-band localisation, arrowheads show spots and/or streaks in the heterozygous cells. Bars, 5 µm.