Figure 6

Topology of the 4q35 region is identical in control and FSHD pluripotent stem cells. Three-Dimension DNA FISH assay within the 4q35 locus in human Induced Pluripotent Stem Cells (hIPSC) generated from control and FSHD1 primary fibroblasts (A, N; respectively - see Table 1). 3D FISH was done using two sets of probes corresponding either to the D4S139 region (red) or different loci along the 4q35 chromosome (green, FAT1, SORBS2, ASCL1 or WWC2; respectively). Representative pictures and 3D reconstruction using IMARIS along with their associated quantifications are presented. We measured the distance of each probe to the nuclear periphery and distance between the probes. For each cell, we evaluated if signals are separated or adjacent (sep, adj; respectively). Scattergram distribution of events and associated Chi-Square test (n = 30 cells per sample, 60 alleles) are shown. Scale is indicated by a white bar (=4 µm). No statistical differences were found between cells. (A) Localization between the D4S139 region and the FAT1 gene. (B) Localization between the D4S139 region and the SORBS2 gene. (C) Localization between the D4S139 region and the ACSL1 gene. (D) Localization between the D4S139 region and the WWC2 gene.