Figure 6.
Boosting NAD+ levels reduces amyloid-like deposits in muscle cells and in vivo in aged mice
(A–C) Representative confocal images of primary human muscle cells from an aged and an IBM donor (scale bar, 50 μm; A and B) or APPSwe-expressing C2C12 myotubes (scale bar, 10 μm; C) treated with NR (3 mM) and stained with the Proteostat fluorescent dye. See also Figures S6A–S6C for quantification.
(D) Microscopy images of TMRM staining of C2C12 APPSwe treated with NR (3 mM; n = 10) and relative quantification. Scale bar, 200 μm.
(E) Representative images and corresponding quantification of immunostainings of A11-positive protein deposits in TA muscles of young or aged male C57BL/6J mice, fed for 8 weeks with chow diet (CD) or chow diet supplemented with NR (400 mg/kg/day; young, n = 12 animals; young+NR, n = 7 animals; old, n = 12 animals; old+NR, n = 11 animals). Scale bar, 50 μm. ∗∗∗p < 0.001 (relative % area).
(F) MSR transcript analysis of forelimbs muscles of aged male mice C57BL/6J following NR treatment (n = 5 animals per group).
(G) Immunoblot of the OXPHOS proteins, MTCO1 and SDHB, from forelimb muscles of the animals in (E) and Figure S4J (n = 3 animals per group).
See STAR methods for further details. Values in the figure are mean ± SEM. ∗p < 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001. Differences for two groups were assessed using two-tailed t tests (95% confidence interval). For all the individual p values, see the Table S6 (Excel data source Figure 6).