Extended Data Fig. 9: Myo-granules that seed amyloid-like fibres are increased in human muscle regeneration and in multisystem proteinopathy. | Nature

Extended Data Fig. 9: Myo-granules that seed amyloid-like fibres are increased in human muscle regeneration and in multisystem proteinopathy.

From: TDP-43 and RNA form amyloid-like myo-granules in regenerating muscle

Extended Data Fig. 9

Related to Figs. 4, 5. a, Representative images of TDP-43 expression (top) and secondary antibody-only control (bottom) in regenerating human skeletal muscle from a patient with necrotizing myopathy. n = 3 independent patient biopsies, each showing similar results. Scale bars, 50 μm. b, Representative tibialis anterior cross-section images of uninjured VCP(A232E) and VCP(WT) mice labelled with EdU after 21 days of EdU treatment in the drinking water to mark division and fusion of muscle stem cells. Laminin identifies myofibres and cells are stained with DAPI to identify nuclei. Arrowheads indicate myofibres with EdU+ centrally located myonuclei. n = 3 mice, each showing similar results. Scale bars, 200 μm and 50 μm (inset). c, Quantification of myofibres with EdU+ centrally located myonuclei in VCP(A232E) and VCP(WT) mice. n = 4 mice, over 1,000 myofibres were quantified per genotype. Data are mean ± s.d. Unpaired, two-tailed Student’s t-test, P = 6.5 × 10−6. d, Representative deconvolution image of A11 and TDP-43 co-localization in a regenerating myofibre from a VCP(A232E) tibialis anterior muscle. n = 3 mice, each showing similar results. Scale bar, 10 μm. e, Secondary antibody-only control of uninjured VCP(A232E) tibialis anterior muscle sections reveals a lack of signal. Nuclei were counterstained with DAPI and myofibres were outlined in white. n = 4 mice, each showing similar results. Scale bar, 25 μm. f, Coomassie-stained recombinant HIS–SUMO–TDP-43 used for thioflavin-T assays resolved by SDS–PAGE. n = 3 biologically independent experiments, each showing similar results. g, Thioflavin-T incorporation reveals thioflavin-T+ amyloid-like fibres for recombinant amyloid-β1–42 and absence of thioflavin-T signal in both the IgG pull-down control and thioflavin-T alone. n = 3 biologically independent experiments, each showing similar results. Scale bars, 10 μm. h, Representative TEM image (zoomed out from Fig. 5e) of thioflavin-T+ (ThT) fibres formed from isolated myo-granules. n = 3 biologically independent experiments. Scale bar, 1 μm.

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