PMC4910231_01
Family pedigree showing the carriers of the (X;22) translocation and X mosaicism
13039_2016_249_Fig1_HTML.jpg
PMC4910231_01_13039_2016_249_Fig1_HTML.jpg
Fig1
Figure 1 shows the family pedigree.
FISH and Cytogenetic analyses of the translocation cariers. a) FISH technique from patient A2 showing the subtelomeric probe of chromosome 22 (in green) in the distal portion of the short arm of the translocated X chromosome, while probe for TUPLE1 gene is located at band q11.21 (in red), remaining on the der(22). b) On the left side the ideograms from normal chromosomes X and 22 show the breakpoints (red arrow) and the translocated segments (shaded in red). On the right side are the schematic ideograms and the GTG banding pattern of the derivative chromosomes
13039_2016_249_Fig2_HTML.jpg
PMC4910231_01_13039_2016_249_Fig2_HTML.jpg
Fig2
ANOS1) mapped to p22.31 remained on the derivative X chromosome and did not translocate to the chromosome 22. Additionally, the subtelomeric probe of chromosome 22 (N85A3) was displayed in the terminal portion of the short arm of the translocated X chromosome, while the TUPLE1 gene (also known as HIRA), mapped to 22q11.21, remained on the chromosome 22 translocated (Fig. 2a). Figure 2b shows a schematic image of the position of the genes investigated by FISH in normal and translocated chromosomes.
Results of X inactivation assay. Electropherograms using the 5meCpG-based RP2/AR repeat biplex PCR assay across the HpaII restriction site in the mother and daughter carrying a balanced X- autosometranslocation. Both mother and daughter are homozygous for the AR marker (allele 233bp). For the informative RP2 marker, the (X;22) translocated chromosome was identified by the maternal segregation pattern of the shared 372bp allele. The pattern of inactivation demonstrates that translocated (X;22) chromosome is active in 99.3% of the blood cells in the mother and in 88.0% of blood cells in the daughter. The boxed numbers correspond to the amplicon length (top) and the areas under the allele peaks (bottom). RFU: Relative fluorescence units
13039_2016_249_Fig3_HTML.jpg
PMC4910231_01_13039_2016_249_Fig3_HTML.jpg
Fig3
Segregation analysis of the RP2 alleles indicated that the X chromosome represented by the 372 bp allele corresponds to the X;22 translocated chromosome, and that it was preferentially active in both mother and daughter (Fig. 3).