A large cohort study confirming that specific haplotype 4A161PAS is exclusively associated with the Chinese FSHD1

Fig. S1. Schematic overview of the 4q and 10q subtelomeric region with essential polymorphic markers. (1) The simple sequence length polymorphism (SSLP) is localized 3.5-kb proximal to D4Z4 array in the 4q35/10q26 region. (2) The D4Z4 repeat array resides in subtelomeric region of chromosomes 4q35 and 10q26 indicated with triangles. Both chromosomes 4q35 and 10q26 contain EcoRI and HindIII enzyme restriction-recognition sites, but the presence of Bln enzyme restriction-recognition sites only exist in 10q-dervied units. (3) The distal variant 4qA or 4qB in subtelomere of chromosomes 4q35 and 10q26, which could be detected by Hind III digestion and hybridized by probe 4qA or 4qB. (4) PLAM sequence adjacent to the terminal D4Z4 repeats in ‘permissive’ haplotype. Fig. S2. Analysis of the polymorphic markers in chromosome 4q35/10q26 region in the control individual. (1) Genomic DNA was digested with EcoRI (lane E) and EcoRI/BlnI (lane B), further separated by pulsed-field gel electrophoresis (PFGE) and then blotted DNA fragments were visualized followed hybridization with probe p13E-11. 4q-derived repeat arrays are resistant to BlnI, whereas 10q-dervied repeat arrays are sensitive to BlnI. The individual carried two 4q-dervied alleles of 76kb and 95.5 kb, and two 10q-dervied alleles of 36 kb and 59.5 kb. (2) DNA with HindIII digestion (lane H) was hybridized with probes 4qA and 4qB. Both 4q-derived alleles (76 and 95.5 kb) were identified as 4qB variants, whereas two 10q-derived alleles (36 and 59.5kb) were identified as 4qA variants. (3) After PFGE, DNA fragments were extracted from gel slices and amplified by the specific the primers, further followed by capillary electrophoresis. The simple sequence length polymorphism (SSLP) fragment analysis showed that both 4q-derived alleles were ran at the peak of 163 bp, in combination with the distal variant 4qB, and the haplotype of chromosome 4q35 in the individual was 4qB163 homozygote. Analysis of 10q-dervied alleles showed that one allele (59.5kb) were ran at the peak of 164 bp, and another allele (36 kb) were ran at the peak of 166 bp. In combination with the distal variant 4qA, the haplotype of chromosome 10q26 in the individual were heterozygosis 10qA164/166. Fig. S3. The proportions of haplotypes distribution in chromosome 4q35. Fig. S4. Comparsion the different D4Z4 repeats of 136 unrelated families among sporadic, familial and uncertain patients. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.