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Figure 5


Fig. 5. Endogenous obscurin localization in NRCs transfected with fragments related to obscurin/Obsl1-myomesin interaction. (A) Overexpression of the myomesin-binding third Ig domains of obscurin (Ob-Ig3) or Obsl1 (Obsl1-Ig3) leads to loss of endogenous obscurin (Ob58-59) from the M-band. Note the strong reduction of obscurin M-band staining with diffuse localization in the transfected cells, visualized by GFP fluorescence of the tagged transfected constructs. Similarly, transfection of the myomesin My4-My5 linker leads to a reduction of M-band-bound obscurin with broad, fuzzy stripes. Overlay and individual channels are shown. (Inset) Twofold-magnified area of the same image showing broad doublet stripes (arrows). (B) Endogenous Obsl1 in NRCs transfected with fragments related to obscurin/Obsl1-myomesin interaction. Overexpression of the myomesin-binding third Ig domains of obscurin (Ob-Ig3) or Obsl1 (Obsl1-Ig3) leads to loss of endogenous Obsl1 from the M-band. Note the reduction of Obsl1 M-band staining with diffuse localization in the transfected cells, visualized by GFP fluorescence of the tagged transfected constructs. Similarly, transfection of the myomesin My4-My5 linker leads to a reduction of M-band-bound Obsl1, with broader, fuzzy stripes (arrows). Scale bars: 10 µm.