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. 2021 Jan 19;34(3):108660. doi: 10.1016/j.celrep.2020.108660

Figure 2.

Figure 2

Accumulation of amyloid-like deposits in aging muscle across species

(A and B) Representative confocal images of primary human muscle cells from young, aged, and IBM donors (scale bar, 50 μm; A; representative images of n = 2 individuals per group) or of control and APPSwe-expressing C2C12 myotubes (scale bar, 10 μm) stained with the Proteostat fluorescent dye (B; n = 3 biological replicates per group).

(C) Microscopy images of tetramethylrhodamine methyl ester (TMRM) staining of C2C12 ev, C2C12 ev treated with trifluoromethoxy carbonylcyanide phenylhydrazonen (FCCP) (10 μM; internal control for membrane depolarization), and C2C12 APPSwe (C2C12 ev + Veh, n = 17; C2C12 ev + FCCP, n = 5; C2C12 APPSwe + Veh, n = 20 biological replicates) and relative quantification. Scale bar, 200 μm. ∗∗∗∗p < 0.0001.

(D) Representative images and corresponding quantification of immunostainings of A11-positive deposits in tibialis anterior (TA) muscles of young (3 months) or aged (24 months) male C57BL/6J mice (n = 12 animals). Scale bar, 50 μm. ∗∗∗p < 0.001 (relative % area).

(E) Dot blot of protein lysates from young (day 1) and aged N2 worms (day 11; n = 3 biological replicates per group) and from GMC101 worms, fed with bacteria containing either an empty vector (e.v.) or the atfs-1 RNAi. Dot blots were stained for the presence of oligomers using the A11 antibody. Relative quantification of the blot intensities is reported in Table S4.

(F) Confocal images of TMRM staining of young (day 1) and aged (day 11) N2 worms (day 1, n = 8; day 11, n = 5 worms) and relative quantification. Scale bar, 100 μm. ∗∗∗p < 0.001.

(G and H) Loss of healthy mitochondrial morphology in SJ4103 worms (G) and muscle fiber structural decay in the RAW1596 (myo-3p::GFP) C. elegans strain (H) between day 1 and day 11 of aging.

(I) A11 dot blotting of N2 (day 6) worms treated with pnc-1 RNAi (n = 2 biological replicates per group).

Relative quantification of the blot intensities is reported in Table S4. See also Figures S1 and S2. For all the individual p values, see Table S6 (Excel data source Figure 2). Values in the figure are mean ± S.E.M.