Chromatin loop domain organization within the 4q35 locus in facioscapulohumeral dystrophy patients versus normal human myoblasts -- Petrov et al. 103 (18): 6982 Data Supplement - HTML Page - index.htslp -- Proceedings of the National Academy of Sciences

Petrov et al. 10.1073/pnas.0511235103.

Supporting Information

Files in this Data Supplement:

Supporting Figure 7
Supporting Table 1
Supporting Figure 8

Fig. 7. FISH on the nuclear halos from the primary myoblasts from normal subjects and FSHD patients. Hybridization of the FRR MAR (red) and the D4Z4 repeats (green) to the nuclear halos from human primary myoblasts from two different unaffected individuals (A and B), and those from two FSHD patients (C and D). The positions of FISH signal corresponding to FRR MAR are indicated by arrows. The first image in each row shows nuclear halos counterstained with DAPI (blue); the second image shows FISH signals from FRR MAR (red); the third image shows FISH signals corresponding to D4Z4 (green); the fourth image shows the merged results of the two previous images; and the fifth image shows paths of chromatin loops (green) relative to the nuclear matrices and halos (blue).

Fig. 8. Visualization of DNA loops from the 4q35 locus. Hybridization of the bacterial artificial chromosome probe, which contains an insert covering the region between 90 and 250 kb relative to the D4Z4 array (green) of the nuclear halos from human primary myoblasts of two different unaffected individuals (A–A'′′ and B–B'′′) and those from two FSHD patients (C–C'′′ and D–D'′′). (A–D) DNA stained with DAPI (blue). (A'–D') The results of hybridization (green). (A'′--D'′) Merged images. (A'′′–D'′′) The paths of chromatin loops (green) relative to the nuclear matrices and halos (blue).

Table 1. Distribution of hybridization signals over nuclear cores or loop halos

Cell type	Probe	No. of nuclei examined	Signals in the nuclear matrix, %	Signals in the halo, %
GM10115A	D4Z4	61	2	98
GM10115A	FRR MAR	55	91	9
Normal myoblasts	D4Z4	51	12	88
Normal myoblasts	FRR MAR	57	89	11
FSHD myoblasts	D4Z4	60	15	85
FSHD myoblasts	FRR MAR	51	67	33

Nuclear halos were hybridized with probes that detect loops or attachment regions as mapped in Fig. 4. Nuclei were examined, and the numbers of signals corresponding to D4Z4 and FRR MAR in nuclear core or loop regions were counted.