Genetic defects are common in myopathies with tubular aggregates

Figures

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  Figure 1. Pedigrees of the Family A and Family B. Black symbols represent affected patients; Black arrow points the index patient of the family; Symbols with slash indicate deceased individuals; Numbers inside the symbols indicate multiple individuals with the same gender; Diamond with a question mark indicates that the number of individuals with unknown gender was unclear.

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  Figure 2. Pathological changes observed in Case 2. Hematoxylin and eosin staining shows variation in fibre size with frequent atrophic fibres and increased connective tissue. Prominent basophilic inclusions are present in many fibres (A); Modified Gomori trichrome staining highlights bright red inclusions (B); The inclusions were stained dark in NADH‐TR (C) and adenylate deaminase preparations (G); The inclusions were immunoreactive for SERCA II (D); Fast myosin (E) and slow myosin (F) immunohistochemistry indicated both type I and type II fibres were affected; Accumulations of double‐walled tubular aggregates were confirmed in electron microscopy (EM) (H). Inclusions of tubular aggregates are indicated by arrows in images A‐C, G and H. Scale bar represents 50 μm in A‐G for histology images; Scale bar in H represents 500 nm in EM

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  Figure 3. Pathological changes observed in Case 3 and Case 5. Case 3 (A‐D): There were numerous subsarcolemmal and internal basophilic inclusions suggesting TAs in muscle fibres stained using hematoxylin and eosin (A), while these inclusions were stained red in the modified Gomori trichrome preparation (B), and were stained as dark in NADH‐TR and mainly in type II fibres (C); In toluidine blue, the inclusions were in dark blue, with small dark dots in fibres representing lipid droplets (D); Case 5 (E‐H): There was increased variation in fibre size and scattered atrophic fibres which were mostly angular or polygonal in outline. Occasional fibres contain basophilic inclusions suggestive of TAs when stained with hematoxylin and eosin (E, F); TAs are stained red in the modified Gomori trichrome preparation (G), and are darkly stained in NADH‐TR (H). Inclusions of TAs are indicated by arrows in all the images. Scale bar represents 25 μm in A, B, D and F‐H; represents 50 μm in C and E.

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  Figure 4. Functional analysis of the variant p. Ala11Thr in the ALG14 gene. (A) Western blot of HEK293 cells with wild type and p. Ala11Thr mutant ALG14. Transfection efficiency was verified by co‐transfection of EGFP. (B) The level of ALG14 expression was the mean value of three experiments, and the error bar represents the standard deviation. EGFP, enhanced green fluorescent protein; HEK, human embryonic kidney 293; WT, wild type.