Supplementary material:
Yonekawa et al. Sialyllactose ameliorates myopathic phenotypes in symptomatic GNE
myopathy mice
Supplementary figure legends
Fig. 1. NeuAc treatment for symptomatic GM mice with treadmill evaluation. (A) NeuAc
treatment for symptomatic mice. GM mice were divided into two groups: NeuAc group (n =
8), NeuAc was given at 1,000 mg/kg/d; Non-treated group (n = 17), acid water was given.
Control littermates were also divided. Two in NeuAc group and 10 mice in non-treated group
died during this study, respectively. (B) Body weight losses of tested mice at the end point
from starting are shown; NeuAc (red circle), non-treated (open circle), and control littermates
(black square). (C) Chronological treadmill motor performance in LM, GM treated with
NeuAc, and non-treated GM mice groups.
Fig. 2. Body weight change of mice during treatment. GM mice with high dose 6’-SL, HD
(upward closed triangles, n = 8); low dose 6’-SL, LD (downward closed triangles, n = 10);
GM mice with NeuAc (closed diamonds, n = 10); non-treated GM, NT (open circles, n = 18);
and control littermates, LM (closed circles, n = 5).
Fig. 3. Spontaneous locomotion activities measured chronologically at ages 50, 65, 72
and 80 weeks. (A) Control littermates (n = 14), (B) Non-treated GM mice (n = 14), (C) GM
mice with high dose 6’-SL (n = 5), (D) GM mice with low dose 6’-SL (n = 7), (E) GM mice
with NeuAc (n = 8). *p < 0.05, **p < 0.01, ns; not significant.
Fig. 4. Immunohistochemical staining for laminin-α2 and Aβ1-42 of gastrocnemius
cryosections. NT, non-treated; HD, high dose of 6’-SL; LD, low dose of 6’-SL.
Fig. 5. Recovery of muscle atrophy and weakness in tibialis anterior muscle after
treatment with 6’-SL. (A) Weight of the isolated tibialis anterior muscle in GM model mice
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treated either with high dose 6’-SL (HD), low dose 6’-SL (LD), or NeuAc as compared to
non-treated GM (NT) mice. (B) Tibialis anterior whole muscle CSA treated either with HD,
LD, or NeuAc as compared to NT mice. (C–F) Contractile properties of the tibialis anterior
muscle as determined by ex vivo measurement of isometric force (Pt) (C), tetanic force (Po)
(D), specific isometric force (Pt/CSA) (E), and specific tetanic force (Po/CSA) (F). Control
littermates (black bars, n = 14); NT mice (white bars, n = 15); GM mice with HD (dark gray
bars, n = 5); GM mice with LD (gray bars, n = 7); GM mice with NeuAc (light gray bars, n =
9). *p < 0.05, **p < 0.01, ns; not significant.
Fig. 6. Histogram of total individual muscle fiber diameters in an each of LM and 4 GM
mice groups. The histograms in affected mice group are shifted to the left as the frequency of
atrophic fibers are increased. (A) Control littermates (n = 14). (B) Non-treated GM mice (n =
15). (C) GM mice with high dose 6’-SL (n = 5). (D) GM mice with low dose 6’-SL (n = 7).
(E) GM mice with NeuAc (n = 9).
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Supplementary table
Table 1 Effect of sialic acid metabolites on liver and kidney function.
AST (IU/L)
ALP (IU/L)
BUN (mg/dl)
LM (n = 5)
25.0 ± 16.2
17.0 ± 16.4
24.6 ± 9.0
NT (n = 15)
167.3 ± 168.2
100.1 ± 78.3
33.7 ± 12.1
HD (n = 5)
121.5 ± 70.2
64.4 ± 68.6
19.4 ± 7.3
LD (n = 7)
141.7 ± 91.1
95.1 ± 46.9
27.5 ± 8.1
NeuAc (n = 9)
79.3 ± 38.2
111.0 ± 72.6
30.7 ± 9.0
The blood samples were collected from 80-week-old LM and GM after treatment. Data are
mean ± S.D. LM, control littermates; NT, non-treated GM mice; HD, GM mice with high
dose 6’-SL; LD, GM mice with low dose 6’-SL; NeuAc, GM mice with NeuAc.
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Supplementary figures
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